AU2254601A - Beta2-adrenergic receptor agonists - Google Patents

Abstract

Disclosed are multibinding compounds which are beta 2 adrenergic receptor agonists and are useful in the treatment and prevention of respiratory diseases such as asthma, bronchitis. They are also useful in the treatment of nervous system injury and premature labor.

Description

WO 01/42193 PCT/USOO/33057 p2-ADRENERGIC RECEPTOR AGONISTS BACKGROUND OF THE INVENTION Field of the Invention This invention relates to novel multibinding compounds (agents) that are p2 adrenergic receptor agonists, partial agonists and pharmaceutical compositions comprising such compounds. Accordingly, the multibinding compounds and 5 pharmaceutical compositions of this invention are useful in the treatment and prevention of respiratory diseases such as asthma, chronic obstructive pulmonary disease and chronic bronchitis. They are also useful in the treatment of nervous system injury and premature labor. 10 References The following publications are cited in this application as superscript numbers: Hardman, J. G., et al. "The Pharmacological Basis of Therapeutics", 15 McGraw-Hill, New York, (1996) 2 Strosberg, A. D. "Structure, Function, and Regulation of Adrenergic Receptors" Protein Sci. 2, 1198-1209 (1993).

WO 01/42193 PCT/USOO/33057 --2- 3 Beck-Sickinger, A. G. "Structure Characterization and Binding Sites of G-Protein-coupled Receptors" DDT, 1, 502-513, (1996). 4 Hein, L. & Kobilka, B. K. "Adrenergic Receptor Signal 5 Transduction and Regulation" Neuropharmacol, 34, 357-366, (1995). 5 Strosberg, A. D. & Pietri-Rouxel, F. "Function, and Regulation of p3-Adrenoceptor" TiPS, 17, 373-381, (1996). 10 6 Barnes, P. J. "Current Therapies for Asthma" CHEST, 111:17S-26S, (1997). Jack, D. A. "A way of Looking at Agonism and Antagonism: 15 Lessons from Salbutamol, Salmeterol and other P Adrenoceptor Agonists" Br. J Clin. Pharmac. 31, 501-514, (1991). 8 Kissei Pharmaceutical Co. Ltd. "2-Amino-1-(4-hydroxy-2-methyl phenyl)propanol derivatives" JP- 10152460 (Publication date June 9, 20 1998). All of the above publications are herein incorporated by reference in their entirety to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference in its entirety. 25 State of the Art A receptor is a biological structure with one or more binding domains that reversibly complexes with one or more ligands, where that complexation has biological consequences. Receptors can exist entirely outside the cell (extracellular 30 receptors), within the cell membrane (but presenting sections of the receptor to the extracellular milieu and cytosol), or entirely within the cell (intracellular receptors). They may also function independently of a cell (e.g., clot formation). Receptors within the cell membrane allow a cell to communicate with the space outside of its boundaries (i.e., signaling) as well as to function in the transport of molecules and 35 ions into and out of the cell. A ligand is a binding partner for a specific receptor or family of receptors. A ligand may be the endogenous ligand for the receptor or alternatively may be a WO 01/42193 PCT/USOO/33057 --3synthetic ligand for the receptor such as a drug, a drug candidate or a pharmacological tool. The super family of seven transmembrane proteins (7-TMs), also called G-protein coupled receptors (GPCRs), represents one of the most significant classes 5 of membrane bound receptors that communicate changes that occur outside of the cell's boundaries to its interior, triggering a cellular response when appropriate. The G-proteins, when activated, affect a wide range of downstream effector systems both positively and negatively (e.g., ion channels, protein kinase cascades, transcription, transmigration of adhesion proteins, and the like). 10 Adrenergic receptors (AR) are members of the G-protein coupled receptors that are composed of a family of three receptor sub-types: P1 (A, B, D) P2 (A, B, c), and (, 2,3)."- These receptors are expressed in tissues of various systems and organs of mammals and the proportions of the a and the p receptors are tissue dependant. For example, tissues of bronchial smooth muscle express largely P2-AR while 15 those of cutaneous blood vessels contain exclusively p-AR subtypes. It has been established that the P2-AR sub-type is involved in respiratory diseases such as such as asthma, chronic bronchitis, nervous system injury, and premature labor'. Currently, a number of drugs e.g., albuterol, formoterol, isoprenolol, or salmeterol having P2-AR agonist activities are being used to treat 20 asthma. However, these drugs have limited utility as they are either non-selective thereby causing adverse side effects such as muscle tremor, tachycardia, palpitations, and restlesness', or have short duration of action and/or slow onset time of action.' Accordingly, there is a need for P2-selective AR agonists that are fast acting and have increased potency and /or longer duration of action. 25 The multibinding compounds of the present invention fulfill this need. SUMMARY OF THE INVENTION This invention is directed to novel multibinding compounds (agents) that are agonists or partial agonists of P2 adrenergic receptor and are therefore useful in the 30 treatment and prevention of respiratory diseases such as asthma, chronic obstructive WO 01/42193 PCT/USOO/33057 --4pulmonary disease, and chronic bronchitis. They are also useful in the treatment of nervous system injury and premature labor. Accordingly, in one of its composition aspects, this invention provides a multibinding compound of Formula (I): 5 (L)p(X)q (I) wherein: p is an integer of from 2 to 10; q is an integer of from I to 20; 10 X is a linker; and L is a ligand wherein: one of the ligands, L, is a compound of formula (a): OH R 2 A' N-W-Ar2 RI (a) wherein: Ar' and Ar 2 are independently selected from the group consisting of aryl, 15 heteroaryl, cycloalkyl, substituted cycloalkyl, and heterocyclyl wherein each of said Ar' and Ar2 substituent optionally links the ligand to a linker; R' is selected from the group consisting of hydrogen, alkyl, and substituted alkyl, or R' is a covalent bond linking the ligand to a linker;

R

2 is selected from the group consisting of hydrogen, alkyl, aralkyl, acyl, 20 substituted alkyl, cycloalkyl, and substituted cycloalkyl, or R is a covalent bond linking the ligand to a linker; W is a covalent bond linking the -NR 2 - group to Ar2, alkylene or substituted alkylene wherein one or more of the carbon atoms in said alkylene or substituted alkylene group is optionally replaced by a substituent selected from the group 25 consisting of -NRa- (where Ra is hydrogen, alkyl, acyl, or a covalent bond linking WO 01/42193 PCT/USOO/33057 --5the ligand to a linker), -0-, -S(O), (where n is an integer of from 0 to 2), -CO-, PRb- (where R' is alkyl), -P(0) 2 -, and -O-P(O)O- and further wherein said alkylene or substituted alkylene group optionally links the ligand to a linker provided that at least one of Ari, Ar 2 , R', R 2 , or W links the ligand to a linker; and 5 the other ligands are independently of each other a compound of formula (b): -Q-Ar 3 (b) 10 wherein: Ar 3 is selected from the group consisting of aryl, heteroaryl, cycloalkyl, substituted cycloalkyl, and heterocyclyl; Q, which links the other ligand to the linker, is selected from the group consisting of a covalent bond, alkylene, and substituted alkylene wherein one or 15 more of the carbon atoms in said alkylene and substituted alkylene is optionally replaced by a substituent selected from the group consisting of -NRa- (where Ra is hydrogen, alkyl, acyl, or a covalent bond linking the ligand to a linker), -0-, -S(O (where n is an integer of from 0 to 2), -CO-, -PR- (where Rb is alkyl), -P(0) 2 -, and O-P(0)0-; and 20 individual isomers, mixtures of isomers and pharmaceutically acceptable salts thereof provided that: (i) when the multibinding compound of Formula (I) is a compound of formula: OHH H OH Ar 1 N-W-- -X---- Ar 3 25 where Ar and Ar 3 are aryl, then W and X both are not alkylene and when W is alkylene-O-, then X is not -0-alkylene; (ii) when the multibinding compound of Formula (I) is a compound of formula: OH 30 Ar N--W-Ar 2 -X-Q-Ar3

R

WO 01/42193 PCT/USOO/33057 --6where Ar' is 4-hydroxy-2-methylphenyl, Ai is aryl, Ar 3 is aryl or heterocyclyl, W is ethylene, Q is a covalent bond, R' is alkyl, then the linker X is not linked to the Ar2 group through an oxygen atom; (iii) when the multibinding compound of Formula (I) is a compound of formula: 5 OH R 2 Ar 3 -Q-X-Ar 1 N---W-Ar2 where Ar' and Ar 3 are aryl, R 2 is hydrogen or benzyl, W is alkylene, Ar2 is aryl or 10 cycloalkyl, Q is a covalent bond, then X is not -alkylene-O-; and (iv) when the multibinding compound of Formula (I) is a compound of formula: OH R 2 1 2 3 Ar 1 N-CH-CH 2 -Ar -X-Q-Ar

CH

3 15 where Ar' is 4-benzyloxy-3-formylamino, R 2 is aralkyl, W is -CH(CH 3

)CH

2 -, Ar2 and Ar 3 are phenyl, Q is a covalent bond, then the linker X is not linked to the Ar2 group through an oxygen atom. More preferably, each linker, X, in the multibinding compound of Formula 20 (I) independently has the formula: -Xa-Z-(Ya-Z)m-Xa_ wherein m is an integer of from 0 to 20; 25 Xa at each separate occurrence is selected from the group consisting of -0-, -S-, -NR-, -C(O)-, -C(0)O-, -OC(O)-, -C(O)NR-, -NRC(O)-, C(S), -C(S)O-, -C(S)NR-, -NRC(S)-, or a covalent bond where R is as defined below; Z at each separate occurrence is selected from the group consisting of alkylene, substituted alkylene, cycloalkylene, substituted cycloalkylene, alkenylene, 30 substituted alkenylene, alkynylene, substituted alkynylene, cycloalkenylene, WO 01/42193 PCT/USOO/33057 --7substituted cycloalkenylene, arylene, heteroarylene, heterocyclene, or a covalent bond; each Ya at each separate occurrence is selected from the group consisting of -O-, -C(O)-, -OC(O)-, -C(O)O-, -NR-, -S(O),,-, -C(O)NR'-, -NR'C(O)-, 5 -NR'C(O)NR'-, -NR'C(S)NR'-, -C(=NR')-NR'-, -NR'-C(=NR')-, -OC(O)-NR'-, -NR'-C(O)-0-, -N=C(Xa)-NR'-, -NR' -C(Xa)=N-,-P(O)(OR')-0-, -O-P(O)(OR')-, -S(O),CR'R"-, -S(O).-NR'-, -NR'-S(O), -, -S-S-, and a covalent bond; where n is 0, 1 or 2; R, R' and R" at each separate occurrence are selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, 10 alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic, and X is as defined above. Preferably, q is less than p in the multibinding compounds of this invention. In still another of its composition aspects, this invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and 15 an effective amount of a multibinding compound of Formula (I): (L)p(X)q (I) wherein: 20 p is an integer of from 2 to 10; q is an integer of from 1 to 20; X is a linker; and L is a ligand wherein: one of the ligands, L, is a compound of formula (a): OH R 2 N-W -Ar2 Ri (a) 25 wherein: WO 01/42193 PCT/USOO/33057 --8- Ar' and Ar 2 are independently selected from the group consisting of aryl, heteroaryl, cycloalkyl, substituted cycloalkyl, and heterocyclyl wherein each of said Ar' and Ar2 substituent optionally links the ligand to a linker; R' is selected from the group consisting of hydrogen, alkyl, and substituted 5 alkyl, or R' is a covalent bond linking the ligand to a linker;

R

2 is selected from the group consisting of hydrogen, alkyl, aralkyl, acyl, substituted alkyl, cycloalkyl, and substituted cycloalkyl, or R2 is a covalent bond linking the ligand to a linker; W is a covalent bond linking the -NR 2 - group to Ar2, alkylene or substituted 10 alkylene wherein one or more of the carbon atoms in said alkylene and substituted alkylene is optionally replaced by a substituent selected from the group consisting of -NRa- (where Ra is hydrogen, alkyl, acyl, or a covalent bond linking the ligand to a linker), -0-, -S(O), (where n is an integer of from 0 to 2), -CO-, -PRb- (where R' is alkyl), -P(0) 2 -, and -O-P(O)O- and further wherein said alkylene or substituted 15 alkylene group optionally links the ligand to a linker provided that at least one of Ar', Ar2, R', R 2 , or W links the ligand to a linker; and the other ligands are independently of each other a compound of formula (b): -Q-Ar 3 20 (b) wherein: Ar is selected from the group consisting of aryl, heteroaryl, cycloalkyl, substituted cycloalkyl, and heterocyclyl; 25 Q, which links the other ligand to the linker, is selected from the group consisting of a covalent bond, alkylene, or a substituted alkylene group wherein one or more of the carbon atoms in said alkylene or substituted alkylene group is optionally replaced by a substituent selected from the group consisting of -NRa_ (where Ra is hydrogen, alkyl, acyl, or a covalent bond linking the ligand to a linker), WO 01/42193 PCT/USOO/33057 --9- -0-, -S(O),- (where n is an integer of from 0 to 2), -CO-, -PR- (where R' is alkyl),

-P(O)

2 -, and -O-P(O)O-; and individual isomers, mixtures of isomers and pharmaceutically acceptable salts thereof provided that: 5 (i) when the multibinding compound of Formula (I) is a compound of formula: OHH H OH Ar 1 N-W--- -X N Ar 3 where Ar and Ar 3 are aryl, then W and X both are not alkylene and when W is 10 alkylene-O-, then X is not -0-alkylene; (ii) when the multibinding compound of Formula (I) is a compound of formula: OHH3 Ar 1 N-W-Ar -X-Q-Ar Ri 15 where Ar' is 4-hydroxy-2-methylphenyl, A? is aryl, Ar 3 is aryl or heterocyclyl, W is ethylene, Q is a covalent bond, R1 is alkyl, then the linker X is not linked to the Ar2 group through an oxygen atom; (iii) when the multibinding compound of Formula (I) is a compound of formula: 20 OH R 2 Ar 3 -Q-X-Ar 1 N-W-Ar where Ar' and Ar 3 are aryl, R2 is hydrogen or benzyl, W is alkylene, Ar is aryl or cycloalkyl, Q is a covalent bond, then X is not -alkylene-O-; and 25 (iv) when the multibinding compound of Formula (I) is a compound of formula: OH R 2 Ar 1 N- H-CH 2 -Ar 2-X-Q-Ar3

CH

3 WO 01/42193 PCT/USOO/33057 -- 10where Ar' is 4-benzyloxy-3-formylamino, R2 is aralkyl, W is -CH(CH 3

)CH

2 -, Ar 2 and Ar3 are phenyl, Q is a covalent bond, then the linker X is not linked to the Ar2 group through an oxygen atom. More preferably, each linker, X, in the multibinding compound of Formula 5 (I) independently has the formula: -Xa-Z-(Ya-Z)m-Xa_ wherein m is an integer of from 0 to 20; 10 Xa at each separate occurrence is selected from the group consisting of -0-, -S-, -NR-, -C(O)-, -C(0)O-, -OC(0)-, -C(0)NR-, -NRC(O)-, C(S), -C(S)O-, -C(S)NR-, -NRC(S)-, or a covalent bond where R is as defined below; Z at each separate occurrence is selected from the group consisting of alkylene, substituted alkylene, cycloalkylene, substituted cycloalkylene, alkenylene, 15 substituted alkenylene, alkynylene, substituted alkynylene, cycloalkenylene, substituted cycloalkenylene, arylene, heteroarylene, heterocyclene, or a covalent bond; each ya at each separate occurrence is selected from the group consisting of -O-, -C(O)-, -OC(0)-, -C(O)O-, -NR-, -S(O),-, -C(O)NR'-, -NR'C(O)-, 20 -NR'C(O)NR'-, -NR'C(S)NR'-, -C(=NR')-NR'-, -NR'-C(=NR')-, -OC(O)-NR'-, -NR'-C(O)-O-, -N=C(Xa)-NR'-, -NR'-C(X")=N-,-P(O)(OR')-O-, -O-P(O)(OR')-, -S(O)nCR'R"-, -S(O)n-NR'-, -NR'-S(O), -, -S-S-, and a covalent bond; where n is 0, 1 or 2; R, R' and R" at each separate occurrence are selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, 25 alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic, and X is as defined above. In still another aspect, this invention provides a method of treating diseases mediated by a P2 adrenergic receptor in a mammal, said method comprising administering to said mammal a therapeutically effective amount of a WO 01/42193 PCT/USOO/33057 --11pharmaceutical composition comprising a pharmaceutically acceptable carrier and a multibinding compound of Formula (I): (L)p(X)q 5 (I) wherein: p is an integer of from 2 to 10; q is an integer of from I to 20; X is a linker; and 10 L is a ligand wherein: one of the ligands, L, is a compound of formula (a): OH R 2 N-W -Ar2 R 4 (a) wherein: Ar and Ar 2 are independently selected from the group consisting of aryl, heteroaryl, cycloalkyl, substituted cycloalkyl, and heterocyclyl wherein each of said 15 Ar' and Ar2 substituent optionally links the ligand to a linker; R' is selected from the group consisting of hydrogen, alkyl, and substituted alkyl, or R 1 is a covalent bond linking the ligand to a linker;

R

2 is selected from the group consisting of hydrogen, alkyl, aralkyl, acyl, substituted alkyl, cycloalkyl, and substituted cycloalkyl, or R2 is a covalent bond 20 linking the ligand to a linker; W is a covalent bond linking the -NP2- group to Ar2, alkylene or substituted alkylene wherein one or more of the carbon atoms in said alkylene and substituted alkylene group is optionally replaced by a substituent selected from the group consisting of -NRa- (where Ra is hydrogen, alkyl, acyl, or a covalent bond linking 25 the ligand to a linker), -0-, -S(0)n (where n is an integer of from 0 to 2), -CO-, - WO 01/42193 PCT/USOO/33057 --12- PRb- (where Rb is alkyl), -P(O) 2 -, and -O-P(O)O- and further wherein said alkylene or substituted alkylene group optionally links the ligand to a linker provided that at least one of Ar', Ar 2 , R', R 2 , or W links the ligand to a linker; and the other ligands are independently of each other a compound of formula 5 (b): -Q-Ar 3 (b) wherein: 10 Ar 3 is selected from the group consisting of aryl, heteroaryl, cycloalkyl, substituted cycloalkyl, and heterocyclyl; Q, which links the other ligand to the linker, is selected from the group consisting of a covalent bond, alkylene, and substituted alkylene wherein one or more of the carbon atoms in said alkylene and substituted alkylene group is 15 optionally replaced by a substituent selected from the group consisting of -NRa_ (where Ra is hydrogen, alkyl, acyl, or a covalent bond linking the ligand to a linker), -0-, -S(O),- (where n is an integer of from 0 to 2), -CO-, -PRb- (where Rb is alkyl),

-P(O)

2 -, and -0-P(0)O-; and individual isomers, mixtures of isomers and pharmaceutically acceptable salts 20 thereof provided that: (i) when the multibinding compound of Formula (I) is a compound of formula: OHH H OH Ar N Ar 3 25 where Ar and Ar 3 are aryl, then W and X both are not alkylene and when W is alkylene-O-, then X is not -0-alkylene; (ii) when the multibinding compound of Formula (I) is a compound of formula: OH A_ 30 Ar , N-W-Ar2-X-Q-Ar3 Ri WO 01/42193 PCT/USOO/33057 --13where Ar is 4-hydroxy-2-methylphenyl, Ar is aryl, Ar 3 is aryl or heterocyclyl, W is ethylene, Q is a covalent bond, R' is alkyl, then the linker X is not linked to the Ar 2 group through an oxygen atom; 5 (iii) when the multibinding compound of Formula (I) is a compound of formula: OH R 2 Ar 3 -Q-X-Ar 1 N-W-Ar 10 where Ar and Ar 3 are aryl, R 2 is hydrogen or benzyl, W is alkylene, Ar2 is aryl or cycloalkyl, Q is a covalent bond, then X is not -alkylene-O-; and (iv) when the multibinding compound of Formula (I) is a compound of formula: OH R 2 1 2 3 15 Ar 1 N-CH-CH 2 -Ar -X-Q-Ar

CH

3 where Ar is 4-benzyloxy-3-formylamino, R 2 is aralkyl, W is -CH(CH 3

)CH

2 -, Ar2 and Ar 3 are phenyl, Q is a covalent bond, then the linker X is not linked to the Ar2 group through an oxygen atom. 20 More preferably, each linker, X, in the multibinding compound of Formula (I) independently has the formula: -Xa-Z-(Ya-Z)m-Xa_ wherein 25 m is an integer of from 0 to 20; Xa at each separate occurrence is selected from the group consisting of -0-, -S-, -NR-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR-, -NRC(O)-, C(S), -C(S)O-, -C(S)NR-, -NRC(S)-, or a covalent bond where R is as defined below; Z at each separate occurrence is selected from the group consisting of 30 alkylene, substituted alkylene, cycloalkylene, substituted cycloalkylene, alkenylene, WO 01/42193 PCT/USOO/33057 --14substituted alkenylene, alkynylene, substituted alkynylene, cycloalkenylene, substituted cycloalkenylene, arylene, heteroarylene, heterocyclene, or a covalent bond; each ya at each separate occurrence is selected from the group consisting of 5 -0-, -C(O)-, -OC(O)-, -C(0)O-, -NR-, -S(O)n-, -C(O)NR'-, -NR'C(O)-, -NR'C(O)NR'-, -NR'C(S)NR'-, -C(=NR')-NR'-, -NR'-C(=NR')-, -OC(O)-NR'-, -NR'-C(0)-O-, -N=C(Xa)-NR'-, -NR'-C(Xa)=N-,-P(O)(OR')-O-, -O-P(O)(OR')-, -S(O)nCR'R"-, -S(O)n-NR'-, -NR'-S(O), -, -S-S-, and a covalent bond; where n is 0, 1 or 2; R, R' and R" at each separate occurrence are selected from the group 10 consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic, and X is as defined above. Preferably p is less than q. In still another aspect, this invention is directed to general synthetic methods 15 for generating large libraries of diverse multimeric compounds which multimeric compounds are candidates for possessing multibinding properties for P2 adrenergic receptor. The diverse multimeric compound libraries provided by this invention are synthesized by combining a linker or linkers with a ligand or ligands to provide for a library of multimeric compounds wherein the linker and ligand each have 20 complementary functional groups permitting covalent linkage. The library of linkers is preferably selected to have diverse properties such as valency, linker length, linker geometry and rigidity, hydrophilicity or hydrophobicity, amphiphilicity, acidity, basicity and polarization. The library of ligands is preferably selected to have diverse attachment points on the same ligand, different 25 functional groups at the same site of otherwise the same ligand, and the like. This invention is also directed to libraries of diverse multimeric compounds which multimeric compounds are candidates for possessing multibinding properties for P2 adrenergic receptor. These libraries are prepared via the methods described above and permit the rapid and efficient evaluation of what molecular constraints 30 impart multibinding properties to a ligand or a class of ligands targeting a receptor.

WO 01/42193 PCT/USOO/33057 --15- Accordingly, in one of its method aspects, this invention is directed to a method for identifying multimeric ligand compounds possessing multibinding properties for P2 adrenergic receptor which method comprises: (a) identifying a ligand or a mixture of ligands wherein each ligand 5 contains at least one reactive functionality; (b) identifying a library of linkers wherein each linker in said library comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand; (c) preparing a multimeric ligand compound library by combining at 10 least two stoichiometric equivalents of the ligand or mixture of ligands identified in (a) with the library of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands; and (d) assaying the multimeric ligand compounds produced in (c) above to 15 identify multimeric ligand compounds possessing multibinding properties for P2 adrenergic receptor. In another of its method aspects, this invention is directed to a method for identifying multimeric ligand compounds possessing multibinding properties for P2 adrenergic receptor which method comprises: 20 (a) identifying a library of ligands wherein each ligand contains at least one reactive functionality; (b) identifying a linker or mixture of linkers wherein each linker comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand; 25 (c) preparing a multimeric ligand compound library by combining at least two stoichiometric equivalents of the library of ligands identified in (a) with the linker or mixture of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands; and WO 01/42193 PCT/USOO/33057 --16- (d) assaying the multimeric ligand compounds produced in (c) above to identify multimeric ligand compounds possessing multibinding properties for P2 adrenergic receptor. The preparation of the multimeric ligand compound library is achieved by 5 either the sequential or concurrent combination of the two or more stoichiometric equivalents of the ligands identified in (a) with the linkers identified in (b). Sequential addition is preferred when a mixture of different ligands is employed to ensure heterodimeric or multimeric compounds are prepared. Concurrent addition of the ligands occurs when at least a portion of the multimer comounds prepared are 10 homomultimeric compounds. The assay protocols recited in (d) can be conducted on the multimeric ligand compound library produced in (c) above, or preferably, each member of the library is isolated by preparative liquid chromatography mass spectrometry (LCMS). In one of its composition aspects, this invention is directed to a library of 15 multimeric ligand compounds which may possess multivalent properties for P2 adrenergic receptor which library is prepared by the method comprising: (a) identifying a ligand or a mixture of ligands wherein each ligand contains at least one reactive functionality; (b) identifying a library of linkers wherein each linker in said library 20 comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand; and (c) preparing a multimeric ligand compound library by combining at least two stoichiometric equivalents of the ligand or mixture of ligands identified in (a) with the library of linkers identified in (b) under conditions wherein the 25 complementary functional groups react to form a covalent linkage between said linker and at least two of said ligands. In another of its composition aspects, this invention is directed to a library of multimeric ligand compounds which may possess multivalent properties for P2 adrenergic receptor which library is prepared by the method comprising: WO 01/42193 PCT/USOO/33057 --17- (a) identifying a library of ligands wherein each ligand contains at least one reactive functionality; (b) identifying a linker or mixture of linkers wherein each linker comprises at least two functional groups having complementary reactivity to at least 5 one of the reactive functional groups of the ligand; and (c) preparing a multimeric ligand compound library by combining at least two stoichiometric equivalents of the library of ligands identified in (a) with the linker or mixture of linkers identified in (b) under conditions wherein the complementary functional groups react to form a covalent linkage between said 10 linker and at least two of said ligands. In a preferred embodiment, the library of linkers employed in either the methods or the library aspects of this invention is selected from the group comprising flexible linkers, rigid linkers, hydrophobic linkers, hydrophilic linkers, linkers of different geometry, acidic linkers, basic linkers, linkers of different 15 polarization and amphiphilic linkers. For example, in one embodiment, each of the linkers in the linker library may comprise linkers of different chain length and/or having different complementary reactive groups. Such linker lengths can preferably range from about 2 to 1 OOA. In another preferred embodiment, the ligand or mixture of ligands is 20 selected to have reactive functionality at different sites on said ligands in order to provide for a range of orientations of said ligand on said multimeric ligand compounds. Such reactive functionality includes, by way of example, carboxylic acids, carboxylic acid halides, carboxyl esters, amines, halides, isocyanates, vinyl unsaturation, ketones, aldehydes, thiols, alcohols, anhydrides, and precursors 25 thereof. It is understood, of course, that the reactive functionality on the ligand is selected to be complementary to at least one of the reactive groups on the linker so that a covalent linkage can be formed between the linker and the ligand. In other embodiments, the multimeric ligand compound is homomeric (i.e., each of the ligands is the same, although it may be attached at different points) or 30 heterodimeric (i.e., at least one of the ligands is different from the other ligands).

WO 01/42193 PCT/USOO/33057 --18- In addition to the combinatorial methods described herein, this invention provides for an interative process for rationally evaluating what molecular constraints impart multibinding properties to a class of multimeric compounds or ligands targeting a receptor. Specifically, this method aspect is directed to a method 5 for identifying multimeric ligand compounds possessing multibinding properties for P2 adrenergic receptor which method comprises: (a) preparing a first collection or iteration of multimeric compounds which is prepared by contacting at least two stoichiometric equivalents of the ligand or mixture of ligands which target a receptor with a linker or mixture of linkers 10 wherein said ligand or mixture of ligands comprises at least one reactive functionality and said linker or mixture of linkers comprises at least two functional groups having complementary reactivity to at least one of the reactive functional groups of the ligand wherein said contacting is conducted under conditions wherein the complementary functional groups react to form a covalent linkage between said 15 linker and at least two of said ligands; (b) assaying said first collection or iteration of multimeric compounds to assess which if any of said multimeric compounds possess multibinding properties for P2 adrenergic receptor; (c) repeating the process of (a) and (b) above until at least one 20 multimeric compound is found to possess multibinding properties for P2 adrenergic receptor; (d) evaluating what molecular constraints imparted multibinding properties for P2 adrenergic receptor to the multimeric compound or compounds found in the first iteration recited in (a)- (c) above; 25 (e) creating a second collection or iteration of multimeric compounds which elaborates upon the particular molecular constraints imparting multibinding properties to the multimeric compound or compounds found in said first iteration; (f) evaluating what molecular constraints imparted enhanced multibinding properties to the multimeric compound or compounds found in the 30 second collection or iteration recited in (e) above; WO 01/42193 PCT/USOO/33057 --19- (g) optionally repeating steps (e) and (f) to further elaborate upon said molecular constraints. Preferably, steps (e) and (f) are repeated at least two times, more preferably at from 2-50 times, even more preferably from 3 to 50 times, and still more 5 preferably at least 5-50 times. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 illustrates examples of multibinding compounds comprising 2 ligands attached in different formats to a linker. 10 FIG. 2 illustrates examples of multibinding compounds comprising 3 ligands attached in different formats to a linker. FIG. 3 illustrates examples of multibinding compounds comprising 4 ligands attached in different formats to a linker. FIG. 4 illustrates examples of multibinding compounds comprising >4 15 ligands attached in different formats to a linker. FIGs. 5-15 illustrate synthesis of compounds of Formula (I). DETAILED DESCRIPTION OF THE INVENTION Definitions 20 This invention is directed to multibinding compounds which are 2 adrenergic receptor agonists, pharmaceutical compositions containing such compounds and methods for treating diseases mediated by P2 adrenergic receptor in mammals. When discussing such compounds, compositions or methods, the following terms have the following meanings unless otherwise indicated. Any 25 undefined terms have their art recognized meanings. The term "alkyl" refers to a monoradical branched or unbranched saturated hydrocarbon chain preferably having from 1 to 40 carbon atoms, more preferably 1 to 10 carbon atoms, and even more preferably 1 to 6 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso 30 butyl, n-hexyl, n-decyl, tetradecyl, and the like.

WO 01/42193 PCT/USOO/33057 --20- The term "substituted alkyl" refers to an alkyl group as defined above, having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted 5 amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, 10 -S0 2 -alkyl, -SO2-substituted alkyl, -S0 2 -aryl and -S0 2 -heteroaryl. This term is exemplified by groups such as hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-aminoethyl, 3-aminopropyl, 2-methylaminoethyl, 3-dimethylaminopropyl, 2-sulfonamidoethyl, 2-carboxyethyl, and the like. The term "alkylene" refers to a diradical of a branched or unbranched 15 saturated hydrocarbon chain, preferably having from 1 to 40 carbon atoms, more preferably 1 to 10 carbon atoms and even more preferably 1 to 6 carbon atoms. This term is exemplified by groups such as methylene (-CH 2 -), ethylene

(-CH

2

CH

2 -), the propylene isomers (e.g., -CH 2

CH

2

CH

2 - and -CH(CH 3

)CH

2 -) and the like. 20 The term "substituted alkylene" refers to an alkylene group, as defined above, having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, 25 hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, SO 2 -alkyl, -S0 2 -substituted alkyl, -SO 2 -aryl and -S0 2 -heteroaryl. Additionally, 30 such substituted alkylene groups include those where 2 substituents on the alkylene WO 01/42193 PCT/USOO/33057 --21group are fused to form one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups-fused to the alkylene group. Preferably such fused groups contain from 1 to 3 fused ring structures. 5 The term "alkaryl" or "aralkyl" refers to the groups -alkylene-aryl and substituted alkylene-aryl where alkylene, substituted alkylene and aryl are defined herein. Such alkaryl groups are exemplified by benzyl, phenethyl and the like. The term"heteroaralkyl" refers to the groups -alkylene-heteroaryl and -substituted alkylene-heteroaryl where alkylene, substituted alkylene and heteroaryl 10 are defined herein. Such heteroaralkyl groups are exemplified by pyridin-3-lmethyl, pyridin-3-ylmethyloxy, and the like. The term "alkoxy" refers to the groups alkyl-O-, alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O-, and alkynyl-O-, where alkyl, alkenyl, cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferred alkoxy groups are alkyl-O- and 15 include, by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like. The term "substituted alkoxy" refers to the groups substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl-O- where substituted alkyl, substituted alkenyl, substituted 20 cycloalkyl, substituted cycloalkenyl and substituted alkynyl are as defined herein. The term "alkenyl" refers to a monoradical of a branched or unbranched unsaturated hydrocarbon group preferably having from 2 to 40 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbon aloms and having at least 1 and preferably from 1-6 sites of vinyl unsaturation. Preferred 25 alkenyl groups include ethenyl (-CH=CH 2 ), n-propenyl (-CH 2

CH=CH

2 ), iso propenyl (-C(CH 3

)=CH

2 ), and the like. The term "substituted alkenyl" refers to an alkenyl group as defined above having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, 30 cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted WO 01/42193 PCT/USOO/33057 --22amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, 5 alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -S0 2 -alkyl, -S0 2 -substituted alkyl, -S0 2 -aryl and -S0 2 -heteroaryl. The term "alkenylene" refers to a diradical of a branched or unbranched unsaturated hydrocarbon group preferably having from 2 to 40 carbon atoms, more preferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms and 10 having at least 1 and preferably from 1-6 sites of vinyl unsaturation. This term is exemplified by groups such as ethenylene (-CH=CH-), the propenylene isomers (e.g., -CH 2 CH=CH-, -C(CH 3 )=CH-, and the like. The term "substituted alkenylene" refers to an alkenylene group as defined above having from 1 to 5 substituents, and preferably from 1 to 3 substituents, 15 selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,.acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, 20 aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -S0 2 -alkyl, -S0 2 -substituted alkyl, -S0 2 -aryl and -S0 2 -heteroaryl. Additionally, such substituted alkenylene groups include those where 2 substituents on the alkenylene group are fused to form one or more cycloalkyl, substituted 25 cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fused to the alkenylene group. The term "alkynyl" refers to a monoradical of an unsaturated hydrocarbon preferably having from 2 to 40 carbon atoms, more preferably 2 to 20 carbon atoms and even more preferably 2 to 6 carbon atoms and having at least 1 and preferably WO 01/42193 PCT/USOO/33057 --23from 1-6 sites of acetylene (triple bond) unsaturation. Preferred alkynyl groups include ethynyl (-C= CH), propargyl (-CH 2 C= CH) and the like. The term "substituted alkynyl" refers to an alkynyl group as defined above having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the 5 group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, 10 heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl,

-SO

2 -alkyl, -S0 2 -substituted alkyl, -SO 2 -aryl, and -S0 2 -heteroaryl. The term "alkynylene" refers to a diradical of an unsaturated hydrocarbon preferably having from 2 to 40 carbon atoms, more preferably 2 to 10 carbon atoms 15 and even more preferably 2 to 6 carbon atoms and having at least 1 and preferably from 1-6 sites of acetylene (triple bond) unsaturation. Preferred alkynylene groups include ethynylene (-C = C-), propargylene (-CH 2 C =C-) and the like. The term "substituted alkynylene" refers to an alkynylene group as defined above having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected 20 from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, 25 heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, S0 2 -alkyl, -SO 2 -substituted alkyl, -SO 2 -aryl and -SO 2 -heteroaryl The term "acyl" refers to the groups HC(O)-, alkyl-C(O)-, substituted alkyl C(O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, cycloalkyl-C(O)-, substituted 30 cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl-C(O)-, WO 01/42193 PCT/USOO/33057 --24heteroaryl-C(O)- and heterocyclic-C(O)- where alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl and heterocyclic are as defined herein. The term "acylamino" or "aminocarbonyl" refers to the group -C(O)NRR 5 where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, heterocyclic or where both R groups are joined to form a heterocyclic group (e.g., morpholino) wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein. The term "sulfonylamino" refers to the group -NRSO 2 Ra where R is 10 hydrogen, alkyl, substituted alkyl, aralkyl, or heteroaralkyl, and R is alkyl, substituted alkyl, amino, or substituted amino wherein alkyl, substituted alkyl, aralkyl, heteroaralkyl and substituted amino are as defined herein. The term "aminoacyl" refers to the group -NRC(O)R where each R is independently hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, 15 amino, substituted amino, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, aryl, heteroaryl and heterocyclic are as defined herein. The term "aminoacyloxy" or "alkoxycarbonylamino" refers to the group -NRC(O)OR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, 20 heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein. The term "acyloxy" refers to the groups alkyl-C(O)O-, substituted alkyl C(O)O-, cycloalkyl-C(O)O-, substituted cycloalkyl-C(O)O-, aryl-C(O)O-, heteroaryl-C(0)O-, and heterocyclic-C(O)O- wherein alkyl, substituted alkyl, 25 cycloalkyl, substituted cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein. The term "aryl" refers to an unsaturated aromatic carbocyclic group of from 6 to 20 carbon atoms having a single ring (e.g., phenyl) or multiple condensed (fused) rings (e.g., naphthyl or anthryl). The aryl group may optionally be fused to 30 a heterocyclic or cycloalkyl group. Preferred aryls include phenyl, naphthyl and the WO 01/42193 PCT/USOO/33057 --25like. Unless otherwise constrained by the definition for the aryl substituent, such aryl groups can optionally be substituted with from 1 to 5 substituents, preferably 1 to 3 substituents, selected from the group consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, 5 substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, sulfonylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, 10 -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -S0 2 -alkyl, -SO 2 substituted alkyl, -S0 2 -aryl, -S0 2 -heteroaryl and trihalomethyl. Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro, trihalomethyl, and thioalkoxy. The term "aryloxy" refers to the group aryl-O- wherein the aryl group is as defined above including optionally substituted aryl groups as also defined above. 15 The term "arylene" refers to the diradical derived from aryl (including substituted aryl) as defined above and is exemplified by 1,2-phenylene, 1,3 phenylene, 1,4-phenylene, 1,2-naphthylene and the like. The term "amino" refers to the group -NH 2 . The term "substituted amino" refers to the group -NRR where each R is 20 independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, acyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic provided that both R's are not hydrogen. The term "carboxyalkyl" or "alkoxycarbonyl" refers to the groups 25 "-C(O)O-alkyl", "-C(O)O-substituted alkyl", "-C(O)O-cycloalkyl", "-C(O)O substituted cycloalkyl", "-C(O)O-alkenyl", "-C(O)O-substituted alkenyl", "-C(O)O-alkynyl" and "-C(O)O-substituted alkynyl" where alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl and substituted alkynyl are as defined herein.

WO 01/42193 PCT/USOO/33057 --26- The term "cycloalkyl" refers to cyclic alkyl groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings, said cycloalkyl group may optionally be fused to an aryl or heteroaryl group. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, 5 cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like. The term "substituted cycloalkyl" refers to cycloalkyl groups having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, 10 substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, 15 heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO 2 -alkyl, -S0 2 -substituted alkyl, -S0 2 -aryl and -S0 2 -heteroaryl. The term "cycloalkenyl" refers to cyclic alkenyl groups of from 4 to 20 carbon atoms having a single cyclic ring and at least one point of internal 20 unsaturation. Examples of suitable cycloalkenyl groups include, for instance, cyclobut-2-enyl, cyclopent-3-enyl, cyclooct-3-enyl and the like. The term "substituted cycloalkenyl" refers to cycloalkenyl groups having from 1 to 5 substituents, and preferably 1 to 3 substituents, selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, 25 substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, WO 01/42193 PCT/USOO/33057 --27heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO-alkyl, -S0 2 -substituted alkyl, -S0 2 -aryl and -S0 2 -heteroaryl. The term "halo" or "halogen" refers to fluoro, chloro, bromo and iodo. 5 The term "heteroaryl" refers to an aromatic group of from 1 to 15 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur within at least one ring (if there is more than one ring). The heteroaryl ring may optionally be fused to a cycloalkyl or heterocyclyl ring. Unless otherwise constrained by the definition for the heteroaryl substituent, such heteroaryl groups can be optionally 10 substituted with 1 to 5 substituents, preferably 1 to 3 substituents, selected from the group consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, 15 carboxylalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -S0 2 -alkyl, -S0 2 -substituted alkyl, -S0 2 -aryl, -S0 2 -heteroaryl and trihalomethyl. Preferred heteroaryl substituents include alkyl, alkoxy, halo, cyano, 20 nitro, trihalomethyl, and thioalkoxy. Such heteroaryl groups can have a single ring (e.g., pyridyl or furyl) or multiple condensed rings (e.g., indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl, pyrrolyl and furyl. The term "heteroaryloxy" refers to the group heteroaryl-O-. The term "heteroarylene" refers to the diradical group derived from 25 heteroaryl (including substituted heteroaryl), as defined above, and is exemplified by the groups 2,6-pyridylene, 2,4-pyridinylene, 1,2-quinolinylene, 1,8 quinolinylene, 1,4-benzofuranylene, 2,5-pyridnylene, 2,5-indolenyl, and the like. The term "cycloalkylene" refers to the diradical group derived from cycloalkyl, as defined above, and is exemplified by the groups 1,6-cyclohexylene, 30 1,3-cyclopentylene, and the like.

WO 01/42193 PCT/USOO/33057 --28- The term "substituted cycloalkylene" refers to the diradical group derived from substituted cycloalkyl, as defined above. The term "cycloalkenylene" refers to the diradical group derived from cycloalkyl, as defined above. 5 The term "substituted cycloalkenylene" refers to the diradical group derived from substituted cycloalkenyl, as defined above. The term "heterocycle" or "heterocyclyl" refers to a monoradical saturated unsaturated group having a single ring or multiple condensed rings, from 1 to 40 carbon atoms and from I to 10 hetero atoms, preferably 1 to 4 heteroatoms, selected 10 from nitrogen, sulfur, phosphorus, and/or oxygen within the ring and further wherein one, two, or three of the ring carbon atoms may optionally be replaced with a carbonyl group (i.e., a keto group). Unless otherwise constrained by the definition for the heterocyclic substituent, such heterocyclic groups can be optionally substituted with 1 to 5, and preferably 1 to 3 substituents, selected from the group 15 consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, 20 heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -S0 2 -alkyl, -S0 2 -substituted alkyl, -SO 2 -aryl and -S0 2 -heteroaryl. Such heterocyclic groups can have a single ring or multiple condensed rings. Preferred heterocyclics include morpholino, piperidinyl, and the like. 25 Examples of heteroaryls and heterocycles include, but are not limited to, pyrrole, thiophene, furan, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, 30 isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, WO 01/42193 PCT/USOO/33057 --29imidazoline, pyrrolidine, piperidine, piperazine, indoline, morpholine, tetrahydrofuranyl, tetrahydrothiophene, and the like as well as N-alkoxy-nitrogen containing heterocycles. The term "heterocyclooxy" refers to the group heterocyclic-O-. 5 The term "thioheterocyclooxy" refers to the group heterocyclic-S-. The term "heterocyclene" refers to the diradical group formed from a heterocycle, as defined herein, and is exemplified by the groups 2,6-morpholino, 2,5-morpholino and the like. The term "oxyacylamino" or "aminocarbonyloxy" refers to the group 10 -OC(O)NRR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein. The term "spiro-attached cycloalkyl group" refers to a cycloalkyl group joined to another ring via one carbon atom common to both rings. 15 The term "thiol" refers to the group -SH. The term "thioalkoxy" or "alkylthio" refers to the group -S-alkyl. The term "substituted thioalkoxy" refers to the group -S-substituted alkyl. The term "thioaryloxy" refers to the group aryl-S- wherein the aryl group is as defined above including optionally substituted aryl groups also defined above. 20 The term "thioheteroaryloxy" refers to the group heteroaryl-S- wherein the heteroaryl group is as defined above including optionally substituted aryl groups as also defined above. As to any of the above groups which contain one or more substituents, it is understood, of course, that such groups do not contain any substitution or 25 substitution patterns which are sterically impractical and/or synthetically non feasible. In addition, the compounds of this invention include all stereochemical isomers arising from the substitution of these compounds. The term "pharmaceutically-acceptable salt" refers to salts which retain the biological effectiveness and properties of the multibinding compounds of this 30 invention and which are not biologically or otherwise undesirable. In many cases, WO 01/42193 PCT/USOO/33057 --30the multibinding compounds of this invention are capable of forming acid and/or base salts by virtue of the presence of amino and/or carboxyl groups or groups similar thereto. Pharmaceutically-acceptable base addition salts can be prepared from 5 inorganic and organic bases. Salts derived from inorganic bases, include by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines, di(substituted alkyl) amines, 10 tri(substituted alkyl) amines, alkenyl amines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines, di(substituted alkenyl) amines, tri(substituted alkenyl) amines, cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines, substituted cycloalkyl amines, disubstituted cycloalkyl amine, trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl) amines, tri(cycloalkenyl) 15 amines, substituted cycloalkenyl amines, disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines, aryl amines, diaryl amines, triaryl amines, heteroaryl amines, diheteroaryl amines, triheteroaryl amines, heterocyclic amines, diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amines where at least two of the substituents on the amine are different and are selected from the 20 group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic, and the like. Also included are amines where the two or three substituents, together with the amino nitrogen, form a heterocyclic or heteroaryl group. Examples of suitable amines include, by way of example only, 25 isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like. It should also be understood that other 30 carboxylic acid derivatives would be useful in the practice of this invention, for WO 01/42193 PCT/USOO/33057 --31example, carboxylic acid amides, including carboxamides, lower alkyl carboxamides, dialkyl carboxamides, and the like. Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include 5 hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, 10 salicylic acid, naphthoic acid, 2-hydroxynaphthoic acid, and the like. The term "pharmaceutically-acceptable cation" refers to the cation of a pharmaceutically-acceptable salt. The term "library" refers to at least 3, preferably from 102 to 10' and more preferably from 102 to 10' multimeric compounds. Preferably, these compounds are 15 prepared as a multiplicity of compounds in a single solution or reaction mixture which permits facile synthesis thereof. In one embodiment, the library of multimeric compounds can be directly assayed for multibinding properties. In another embodiment, each member of the library of multimeric compounds is first isolated and, optionally, characterized. This member is then assayed for 20 multibinding properties. The term "collection" refers to a set of multimeric compounds which are prepared either sequentially or concurrently (e.g., combinatorially). The collection comprises at least 2 members; preferably from 2 to 10' members and still more preferably from 10 to 10' members. 25 The term "multimeric compound" refers to compounds comprising from 2 to 10 ligands covalently connected through at least one linker which compounds may or may not possess multibinding properties (as defined herein). The term "pseudohalide" refers to functional groups which react in displacement reactions in a manner similar to a halogen. Such functional groups 30 include, by way of example, mesyl, tosyl, azido and cyano groups.

WO 01/42193 PCT/USOO/33057 --32- The term "protecting group" or "blocking group" refers to any group which when bound to one or more hydroxyl, thiol, amino or carboxyl groups of the compounds (including intermediates thereof) prevents reactions from occurring at these groups and which protecting group can be removed by conventional chemical 5 or enzymatic steps to reestablish the hydroxyl, thiol, amino or carboxyl group (See., T.W. Greene and P.G.H. Wuts, "Protective Groups in Organic Synthesis", 2 nd Ed.). The particular removable blocking group employed is not critical and preferred removable hydroxyl blocking groups include conventional substituents such as allyl, benzyl, acetyl, chloroacetyl, thiobenzyl, benzylidine, phenacyl, t-butyl 10 diphenylsilyl and any other group that can be introduced chemically onto a hydroxyl functionality and later selectively removed either by chemical or enzymatic methods in mild conditions compatible with the nature of the product. Preferred removable thiol blocking groups include disulfide groups, acyl groups, benzyl groups, and the like. 15 Preferred removable amino blocking groups include conventional substituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ), fluorenylmethoxy-carbonyl (FMOC), allyloxycarbonyl (ALOC), and the like which can be removed by conventional conditions compatible with the nature of the product. 20 Preferred carboxyl protecting groups include esters such as methyl, ethyl, propyl, t-butyl etc. which can be removed by mild conditions compatible with the nature of the product. The term "optional" or "optionally" means that the subsequently described event, circumstance or substituent may or may not occur, and that the description 25 includes instances where said event or circumstance occurs and instances where it does not. The term "ligand" or " ligands" as used herein denotes a compound that is a binding partner for a P2 adrenergic receptor and is bound thereto by complementarity. Preferred ligands are those that are either P2 adrenergic receptor 30 agonists, partial agonists, or antagonists. The specific region or regions of the WO 01/42193 PCT/USOO/33057 --33ligand that is (are) recognized by the receptor is designated as the "ligand domain". A ligand may be either capable of binding to the receptor by itself, or may require the presence of one or more non-ligand components for binding (e.g., Ca+ 2 , Mg+ 2 or a water molecule is required for the binding of a ligand to various ligand binding 5 sites). Examples of ligands useful in this invention are described herein. Those skilled in the art will appreciate that portions of the ligand structure that are not essential for specific molecular recognition and binding activity may be varied substantially, replaced or substituted with unrelated structures (for example, with ancillary groups as defined below) and, in some cases, omitted entirely without 10 affecting the binding interaction. The primary requirement for a ligand is that it has a ligand domain as defined above. It is understood that the term ligand is not intended to be limited to compounds known to be useful in binding to P2 adrenergic receptor (e.g., known drugs). Those skilled in the art will understand that the term ligand can equally apply to a molecule that is not normally associated with P2 15 adrenergic receptor binding properties. In addition, it should be noted that ligands that exhibit marginal activity or lack useful activity as monomers can be highly active as multivalent compounds because of the benefits conferred by multivalency. The term "ligand" or " ligands" as used herein is intended to include the 20 racemic forms of the ligands as well as individual enantiomers and diasteromers and non-racemic mixtures thereof. The term "multibinding compound or agent" refers to a compound that is capable of multivalency, as defined below, and which has 2-10 ligands covalently bound to one or more linkers. In all cases, each ligand and linker in the 25 multibinding compound is independently selected such that the multibinding compound includes both symmetric compounds (i.e., where each ligand as well as each linker is identical) and asymmetric compounds (i.e., where at least one of the ligands is different from the other ligand(s) and/or at least one linker is different from the other linker(s)). Multibinding compounds provide a biological and/or 30 therapeutic effect greater than the aggregate of unlinked ligands equivalent thereto WO 01/42193 PCT/USOO/33057 --34which are made available for binding. That is to say that the biological and/or therapeutic effect of the ligands attached to the multibinding compound is greater than that achieved by the same amount of unlinked ligands made available for binding to the ligand binding sites (receptors). The phrase "increased biological or 5 therapeutic effect" includes, for example: increased affinity, increased selectivity for target, increased specificity for target, increased potency, increased efficacy, decreased toxicity, improved duration of activity or action, increased ability to kill cells such as fungal pathogens, cancer cells, etc., decreased side effects, increased therapeutic index, improved bioavailibity, improved pharmacokinetics, improved 10 activity spectrum, and the like. The multibinding compounds of this invention will exhibit at least one and preferably more than one of the above-mentioned affects. The term "univalency" as used herein refers to a single binding interaction between one ligand as defined herein with one ligand binding site as defined herein. It should be noted that a compound having multiple copies of a ligand (or ligands) 15 exhibit univalency when only one ligand is interacting with a ligand binding site. Examples of univalent interactions are depicted below. The term "multivalency" as used herein refers to the concurrent binding of from 2 to 10 linked ligands (which may be the same or different) and two or more 20 corresponding receptors (ligand binding sites) which may be the same or different. For example, two ligands connected through a linker that bind concurrently to two ligand binding sites would be considered as bivalency; three ligands thus connected would be an example of trivalency. An example of trivalent binding, illustrating a multibinding compound bearing three ligands versus a monovalent 25 binding interaction, is shown below: WO 01/42193 PCT/USOO/33057 --35univalent interaction 5 trivalent interaction It should be understood that not all compounds that contain multiple copies of a ligand attached to a linker or to linkers necessarily exhibit the phenomena of 10 multivalency, i.e., that the biological and/or therapeutic effect of the multibinding agent is greater than the sum of the aggregate of unlinked ligands made available for binding to the ligand binding site (receptor). For multivalency to occur, the ligands that are connected by a linker or linkers have to be presented to their ligand binding sites by the linker(s) in a specific manner in order to bring about the desired 15 ligand-orienting result, and thus produce a multibinding event. Furthermore, the multibinding compound of the present invention can be composed of ligands that are all P2 adrenergic receptor agonists, partial agonists, or it can be composed of ligands that are selected from P2 adrenergic receptor agonists and antagonists provided that the multibinding compounds exhibits an overall P2 20 adrenergic receptor agonistic or partial agonistic activity. A multibinding compound that exhibits partial agonist activity at adrenoceptors may provide advantages over a compound that exhibits full agonism. Partial agonism may result in a reduction of the rate of receptor desensitization, receptor recycling, or receptor expression in mammalian tissue. This may result in increased therapeutic benefits WO 01/42193 PCT/USOO/33057 --36from using such an agonist versus a compound which behaves as a full agonist for the chronic treatment of pathological conditions or diseases. A multibinding compound may also, or separately, act as a tissue-selective partial agonist. For example, a multibinding compound with P2 adrenoceptor agonist activity may 5 exhibit a full maximal response in relaxing smooth muscle cells in bronchial tissue but exhibit a partial maximal response to adrenoceptor agonism in cardiac tissue. Thus, a multibinding compound that is a tissue-selective partial agonist may provide a lower incidence of undesired side effects such as positive chronotropism and increases in cardiac output. 10 The term "potency" refers to the minimum concentration at which a ligand is able to achieve a desirable biological or therapeutic effect. The potency of a ligand is typically proportional to its affinity for its ligand binding site. In some cases, the potency may be non-linearly correlated with its affinity. In comparing the potency of two drugs, e.g., a multibinding agent and the aggregate of its unlinked 15 ligand, the dose-response curve of each is determined under identical test conditions (e.g., in an in vitro or in vivo assay, in an appropriate animal model such a human patient). The finding that the multibinding agent produces an equivalent biological or therapeutic effect at a lower concentration than the aggregate unlinked ligand is indicative of enhanced potency. 20 The term "selectivity" or "specificity" is a measure of the binding preferences of a ligand for different ligand binding sites (receptors). The selectivity of a ligand with respect to its target ligand binding site relative to another ligand binding site is given by the ratio of the respective values of Kd (i.e., the dissociation constants for each ligand-receptor complex) or, in cases where a biological effect is 25 observed below the Kd , the ratio of the respective EC 50 's (i.e., the concentrations that produce 50% of the maximum response for the ligand interacting with the two distinct ligand binding sites (receptors)). The term "ligand binding site" denotes the site on the p-adrenergic receptor that recognizes a ligand domain and provides a binding partner for the ligand. The 30 ligand binding site may be defined by monomeric or multimeric structures. This WO 01/42193 PCT/USOO/33057 --37interaction may be capable of producing a unique biological effect, for example, agonism, antagonism, and modulatory effects or it may maintain an ongoing biological event, and the like. It should be recognized that the ligand binding sites of the receptor that 5 participate in biological multivalent binding interactions are constrained to varying degrees by their intra- and inter-molecular associations. For example, ligand binding sites may be covalently joined to a single structure, noncovalently associated in a multimeric structure, embedded in a membrane or polymeric matrix, and so on and therefore have less translational and rotational freedom than if the 10 same structures were present as monomers in solution. The terms "agonism", "partial agonism", and "antagonism" are well known in the art. The term "modulatory effect" refers to the ability of the ligand to change the activity of an agonist or antagonist through binding to a ligand binding site. The term "inert organic solvent" or "inert solvent" means a solvent which is 15 inert under the conditions of the reaction being described in conjunction therewith including, by way of example only, benzene, toluene, acetonitrile, tetrahydrofuran, dimethylformamide, chloroform, methylene chloride, diethyl ether, ethyl acetate, acetone, methylethyl ketone, methanol, ethanol, propanol, isopropanol, t-butanol, dioxane, pyridine, and the like. Unless specified to the contrary, the solvents used 20 in the reactions described herein are inert solvents. The term "treatment" refers to any treatment of a pathologic condition in a mammal, particularly a human, and includes: (i) preventing the pathologic condition from occurring in a subject which may be predisposed to the condition but has not yet been diagnosed with the 25 condition and, accordingly, the treatment constitutes prophylactic treatment for the disease condition; (ii) inhibiting the pathologic condition, i.e., arresting its development; (iii) relieving the pathologic condition, i.e., causing regression of the pathologic condition; or 30 (iv) relieving the conditions mediated by the pathologic condition.

WO 01/42193 PCT/USOO/33057 --38- The term "pathologic condition which is modulated by treatment with a ligand" covers all disease states (i.e., pathologic conditions) which are generally acknowledged in the art to be usefully treated with a ligand for the P2-adrenergic receptor in general, and those disease states which have been found to be usefully 5 treated by a specific multibinding compound of our invention. Such disease states include, by way of example only, the treatment of a mammal afflicted with asthma, chronic bronchitis, chronic pulmonary obstructive disease, and the like. The term "therapeutically effective amount" refers to that amount of multibinding compound which is sufficient to effect treatment, as defined above, 10 when administered to a mammal in need of such treatment. The therapeutically effective amount will vary depending upon the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art. 15 The term "linker", identified where appropriate by the symbol 'X', refers to a group or groups that covalently attaches from 2 to 10 ligands (as identified above) in a manner that provides for a compound capable of multivalency. Among other features, the linker is a ligand-orienting entity that permits attachment of at least two copies of a ligand (which may be the same or different) thereto. Additionally, 20 the linker can be either a chiral or achiral molecule. In some cases, the linker maybe a covalent bond that attaches the ligands in a manner that provides for a compound capable of multivalency. Additionally, in some cases, the linker may itself be biologically active. The term "linker" does not, however, extend to cover solid inert supports such as beads, glass particles, fibers, and the like. But it is 25 understood that the multibinding compounds of this invention can be attached to a solid support if desired. For example, such attachment to solid supports can be made for use in separation and purification processes and similar applications. The extent to which multivalent binding is realized depends upon the efficiency with which the linker or linkers that joins the ligands presents these 30 ligands to the array of available ligand binding sites. Beyond presenting these WO 01/42193 PCT/USOO/33057 --39ligands for multivalent interactions with ligand binding sites, the linker or linkers spatially constrains these interactions to occur within dimensions defined by the linker or linkers. Thus, the structural features of the linker (valency, geometry, orientation, size, flexibility, chemical composition, etc.) are features of 5 multibinding agents that play an important role in determining their activities. The linkers used in this invention are selected to allow multivalent binding of ligands to the ligand binding sites of a P2 adrenergic receptor, whether such sites are located interiorly, both interiorly and on the periphery of the receptor structure, or at any intermediate position thereof. 10 Representative Compounds of Formula (I): I. Representative multibinding compounds of Formula (I) wherein p is 2, q is 1, Arl is 4-hydroxy-3-hydroxymethylphenyl, Ai is 1,4 phenylene, R' and R 2 are hydrogen, X, W, Q, and Ar 3 are as defined in Table A 15 below are: O H X-Q-Ar3 * N' W HO 20 HO Table A Cpd. Stereo- W X -Q-Ar 3 (** = stereochem) # chem. at *C 1A (RS) -(CH 2

)

2 - bond -NH-CH** CH(OH)phenyl **(S) 25 2A (RS) -(CH 2

)

2 - bond -NH-CH 2 ** CH(OH)phenyl ** = (R) 3A (RS) -(CH 2

)

2 - bond -NH-CH 2 r** CH(OH)phenyl ** = (RS) 4A (RS) -(CH 2

)

2 - bond -NH-CH 2 r** CH(OH)-(4-hydroxy-3 hydroxy-methyl)phenyl ** = (RS) WO 01/42193 PCT/USOO/33057 --40- 5A (RS) -(CH 2

)

6 0- bond -(CH 2

)

3 -0 -(CH 2

),-NH-CH

2 -** CH(OH)-(4-hydroxy-3 hydroxyethyl)phenyl ** = ( RS) 6A (RS) -CH2- bond -NH-CH 2 -** CH(OH)-(4-hydroxy-3 hydroxy-methyl)phenyl ** = (RS) 7A (R) -(CH2)2- bond -NH-CH 2 -** CH(OH)phenyl ** (S) 8A (R) -(CH2)2- bond -NH-CH 2 -** CH(OH)phenyl ** = (R) 5 9A (RS) -(CH 2

)

6 -0- bond -O-(CH 2 )-O-[4-(3-hydroxypropyl)]

(CH

2

)

3 phenyl 1OA (RS) -CH 2 *CH(OH)- bond -O-(CH 2 )-** CIH(OH)-(CH 2

)-NH-CH

2 CH 2 -O- * =(RS) ** CH(OH)-(4-hydroxy-3-hydroxy methyl)phenyl ** = (RS) 11A (RS) -(CH 2

)

2 - bond -NH-CH 2 -** CH(OH)-O-naphth-1-yl ** =(RS) 10 II. Representative multibinding compounds of Formula (I) wherein p is 2, q is 1, Ar' is 4-hydroxy-3-hydroxymethylphenyl, Ar is 1,4-phenylene, R' and R 2 are hydrogen, X, W, Q, and Ar 3 , are as defined Table B below are: OH H X-Q-Ar 3 15 * N' HO HO Table B Cpd. Stereo- X -Ar' 20 # chem. at *C

W

WO 01/42193 PCT/USOO/33057 --41- 1B (RS) bond -O-(p-C6H4)-NH-CHr** bond 4-hydroxy-3 CH(OH)- ** = (RS) hydroxymethyl phenyl 2B (RS) bond -0- bond 4-aminophenyl 3B (RS) -(CH 2

)

6 - -O-(CH 2

)

1 w-O-(p-C 6

H

4 )- bond 4-hydroxy-3 0- (CH 2

)

3

-O-(CH

2

)

6 -NH- hydroxy

(CH

2

)

3 - CH 2 ** CH(OH)- methylphenyl ** = (RS) stereochem. 4B (RS) -(CH 2

)

6 - -o -(CH 2

)

6 -0-(p-C 6

H

4 )- bond 4-hydroxy-3 0- (CH 2

)

3

-O-(CH

2

)

5 -NH- hydroxy

(CH

2

)

3 - CH 2 ** CH(OH)- methylphenyl ** = (RS) stereochem. 5 5B (RS) -(CH 2

)

2 - -O-(CH 2

)

4 - bond phenyl III. Representative multibinding compounds of Formula (I) wherein p is 2, q is 1, Ar' is 4-hydroxy-3-hydroxy-methylphenyl, R and R 2 are hydrogen, Ar 3 is (4 10 hydroxy-3-hydroxymethyl)phenyl, X, W, Q, and A are as defined in Table C below are: OH H N. N Ar-X ' 15 HO OH HO OH Table C Cpd. Stereochem. W X Ar 2 Q # at *C 20 iC (RS) bond bond trans-1,4- -NH-CH 2 ** CH(OH) cyclohexane **=(RS) 2C (RS) -CH 2 - bond 1,3- -CHrNH-CHr** cyclohexane CH(OH)- ** = (RS) WO 01/42193 PCT/USOO/33057 --42- 3C (RS) -(CH 2

)

3 - bond 1,4-piperazine -(CH 2

)

3

NH-CH

2 ** CH(OH)- ** = (RS) 4C (RS) bond bond p-menthane -NH-CH 2 r** CH(OH) ** = (RS) 5C (RS) bond bond 1,2-phenylene -CH 2

NH-CH

2 **

CH(OH)

** = (RS) 5 IV. Representative multibinding compounds of Formula (I) wherein p is 2, q is 1, Ar' and Ar 3 are 4-hydroxy-3-hydroxymethylphenyl, R' and R 2 are hydrogen, Q is a bond, and W, Ar 2 , and X are as defined in Table D below are: OH H N'W . Ar HO OH HO OH Table D Cpd. Stereochem. W Ar 2 X 10 # at *C ID (RS) bond 1,4- -(CH 2 )-(p-C 6

HO)-NH-CH

2 cyclohexane ** CH(OH) ** = (RS) stereochem. V. Representative multibinding compounds of Formula (I) wherein p is 2, q is 15 1, R' and R 2 are hydrogen, W is -(CH 2

)

2 -, Ar2 is 1,4-phenylene, -Q-Ar, is [2 hydroxy-2-phenyl]ethylamino, X is a bond and Ar is as defined below are as shown in Table E below: OHH Ar 1 N N H O H WO 01/42193 PCT/USOO/33057 --43- Table E Cpd. # Ar Stereochem. Stereoche at *C m. at **C 1E phenyl (RS) (RS) 5 2E phenyl (R) (S) 3E phenyl (R) (R) 4E 4-amino-3,5-dichlorophenyl (RS) (RS) 5E 4-amino-3,5-dichlorophenyl (R) (R) 6E 4-amino-3,5-dichlorophenyl (S) (S) 10 7E 4-amino-3,5-dichlorophenyl (R) (S) 8E 4-amino-3,5-dichlorophenyl (S) (R) 9E 3-formyl-amino-4-hydroxyphenyl (RS) (RS) 1OE 3-formyl-amino-4-hydroxyphenyl (R) (R) 11E 3-formyl-amino-4-hydroxyphenyl (S) (S) 15 12E 3-formyl-amino-4-hydroxyphenyl (R) (S) 13E 3-formyl-amino-4-hydroxyphenyl (S) (R) VI. Miscellanous compounds: OHH N\/ HO0 HO HN HO 20 HO OH WO 01/42193 PCT/USOO/33057 --44- OH N OH OH N H OH OH N OH HO OH HO PREFERRED EMBODIMENTS 5 While the broadest definition of this invention is set forth in the Summary of the Invention, certain compounds of Formula (I) are preferred. (A) A preferred group is a multibinding compound of Formula (II): OHH N-W-A--X-Q-AP (II) where *C has RS, R, or S stereochemistry; 10 Within this group (A) a more preferred group of compounds is that wherein: (i) Arl is aryl, more preferably Ar is a phenyl ring of formula (c): WO 01/42193 PCT/USOO/33057 --45- R 4 R I R 6 (c) wherein:

R

4 is hydrogen, alkyl, halo, or alkoxy, preferably hydrogen, methyl, fluoro, chloro, or methoxy; 5 R 5 is hydrogen, hydroxy, halo, halo, amino, or -NHSO 2 Ra where Ra is alkyl, preferably hydrogen, hydroxy, fluoro, chloro, amino, or -NHSO 2

CH

3 ; and R' is hydrogen, halo, hydroxy, alkoxy, substituted alkyl, sulfonylamino, aminoacyl, or acylamino; preferably hydrogen, chloro, fluoro, hydroxy, methoxy, hydroxymethyl, -CH 2

SO

2

CH

3 , -NHSO2CH 3 , -NHCHO, -CONH 2 , or -NHCONH 2 ; 10 (ii) Ar' is heteroaryl, more preferably Ar' is 2,8-dihydroxyquinolin-5-yl or 3 bromoisoxazol-5-yl; or (iii) Ar' is heterocyclyl, more preferably Ar' is heterocyclyl fused to an aryl ring, most preferably 6-fluorochroman-2-yl; W is a bond linking the -NR2- group to Ar2, alkylene, or a substituted 15 alkylene group wherein one or more of the carbon atoms in the alkylene and the substituted alkylene group is optionally replaced by-O-, preferably a covalent bond, methylene, ethylene, propylene, -(CH2) 6 -0-(CH2) 3

-,-(CH

2

)

6 -O-, or CH 2

CH(OH)CH

2 -0-; and Ar2 is phenyl wherein the W and the X groups are attached at the 1,2-, 1,3-, 20 and 1,4- positions of the phenyl ring; cyclohexyl optionally substituted with methyl and wherein the W and the X groups are attached at the 1,3- and 1,4- positions of the cyclohexyl ring; or piperazine wherein the W and the X groups are attached at the 1,4- positions of the piperazine ring, preferably 1,4-phenylene.

WO 01/42193 PCT/USOO/33057 --46- Within the above more preferred groups, even more preferred groups of compounds are wherein: (a) X is -0-, -0-alkylene, -0-(arylene)-NH-(substituted alkylene)-, -O-(alkylene)-O-(arylene)-(alkylene)-O-(alkylene)- NH-(substituted alkylene)-, 5 -0-(alkylene)-0-(arylene)-, or -(alkylene)-(cycloalkylene)-NH-(substituted alkylene)-, preferably -0-(CH 2

)

4 -; -CH 2 -(1,4-cyclohexyl)-NH-CH 2 -CH(OH)-; -0-(1,4-phenylene)-NH-CH 2 -CH(OH)-; -0-(CH 2 )w0-0-(1,4-phenylene)-(CH 2

)

3 -0 (CH2) 6

-NH-CH

2 -CH(OH)-; -0-(CH 2

)

6 -0-(1,4-phenylene)-(CH 2

)

3 -0-(CH 2

)

5

-NH

CH

2 -CH(OH)-; or -0-(CH 2

)

6 -0-(1,4-phenylene)-; and 10 Q is a covalent bond; or (b) X is a bond; and Q is a substituted alkylene group wherein one or more of the carbon atoms in said substituted alkylene group is optionally replaced by a heteroatom selected from the group consisting of -NRa- (where Ra is hydrogen, alkyl, or acyl) and -0-, 15 preferably -NH-CH 2 **CH(OH)-; -NH-CHr**CH(OH)-CHrO-; -NH

**CH(CH

2 OH)-; -CH 2

NH-CH

2 -**CH(OH)-; -C(CH3)2-NH-CH 2 -**CH(OH)-; (CH 2

)

3

NH-CH

2 r**CH(OH)-; -(CH2)rO-(CH2)6-NH-CHr**CH(OH)-; -(CH2) 2 NH-CH 2 -**CH(OH)-; -O-(CH 2 )-**CH(OH)-CHrNH-CH**CH(OH)-; or -NH CHr**CH(OH)-CH 2 -0-; more preferably -NH-CH 2 r**CH(OH)-; -NH 20 **CH(CH 2 OH)-; -(CH2)rO-(CH2)6-NH-CHr**CH(OH)-; or -NH-CH 2

**CH(OH)

CH

2 -0- (where ** is RS, R or S stereochemistry), most preferably -NH

**CH(CH

2 OH)- where ** is RS, R or S stereochemistry; Within the above preferred, more preferred group of compounds, a 25 particularly preferred group of compounds is that wherein: (i) Ar 3 is same as Ar as defined in preferred embodiments (A)(i)-(iii) above. Another particularly preferred group of compounds is that wherein: (ii) Ar 3 is a phenyl ring of formula (d): WO 01/42193 PCT/USOO/33057 --47- R 7 R (d) wherein:

R

7 is hydrogen, alkyl, alkenyl, substituted alkyl, halo, alkoxy, substituted alkoxy, hydroxy, aminoacyl, or heteroaryl, preferably hydrogen, methyl, propen-2 5 yl, fluoro, chloro, methoxy, -OCH 2

CO

2 Me, -OCON(CH3)2, hydroxy, -CH2CONH 2 , NHCOCH 3 , -NHCHO, imidazol-1-yl, or 1-methyl-4-trifluoromethylimidazol-2-yl; and R' is hydrogen, halo, alkoxy, substituted alkoxy, or acylamino, preferably hydrogen, fluoro, chloro, methoxy, -OCH 2

CO

2 Me, -OCON(CH 3

)

2 , -NHCHO, or 10 -CONH2 (iii) Yet another particularly preferred group of compounds is that wherein: Ar 3 is naphthyl, pyridyl, benzimidazol-1-yl, indolyl, 2-cyanoindolyl, carbazolyl, 4-methylindanyl, 5-(CH 3 CO2CH 2 0-)-1,2,3,4-tetrahydronaphthyl, 1H-2 oxoindole, 2,3,4-trihydrothianaphthalene, 4-hydroxy-2-benzothiazolinone, or 4 15 oxo-2,3-dihydrothianapthalene. Within the above preferred, more preferred, and particularly preferred groups, even more particularly preferred group is that wherein: Ar 1 is phenyl, 4-hydroxyphenyl, 3,4-dihydroxyphenyl, 3,4-dichlorophenyl, 20 3,5-dihydroxyphenyl, 2-chloro-3,4-dihydroxyphenyl, 2-fluoro-3,4-dihydroxyphenyl, 2-chloro-3,5-dihydroxyphenyl, 2-fluoro-3,5-dihydroxyphenyl, 4-hydroxy-3 methoxyphenyl, 4-hydroxy-3-hydroxymethylphenyl, 4-hydroxy-3-(HCONH )phenyl, 4-hydroxy-3-(NH 2 CO-)phenyl, 3-chlorophenyl, 2,5-dimethoxyphenyl, 4

(CH

3

SO

2 NH-)-phenyl, 4-hydroxy-3-(CH 3

SO

2

CH

2 -)phenyl, 4-hydroxy-3- WO 01/42193 PCT/USOO/33057 --48-

(CH

3

SO

2 NH-)phenyl, 4-hydroxy-3-(NH 2 CONH-)phenyl, 3,5-dichloro-4 aminophenyl, HO or HN 0 preferably 4-hydroxy-3-hydroxymethylphenyl, 4-hydroxy-3-(HCONH-)phenyl, 3,5 dichloro-4-aminophenyl, or HO HN 0 5 most preferably 4-hydroxy-3-(HCONH-)phenyl or 3,5-dichloro-4-aminophenyl; and Ar 3 is: 10 15 WO 01/42193 PCT/USOO/33057 --49- CI x N OCH 2

CO

2 Me (X H; OMe, OH) Me 0

OCH

2

CO

2 Me OCH 2

CO

2 Me ' OCH 2

CO

2 me J OMe K OH I-V-I

CONH

2 - ~ HN CH 2

CONH

2 x (X=H; CN) -r N elMe N\ N NN - ? , \NH

CF

3 s cl OMe X) NHNH (X=0; CH 2 ) lPN

NH

2 OHN

OCON(CH

3

)

2 NH2 OHOH ci CH 2 0H OH NH OCON(CH 3

)

2 ~: x OH N. OH - or OH NHCHOOHH [Y OH, Me, -CONH 2 1 (Z C1 or H) (X C1 or F) (X C1 or F) NHS0 2 Me WO 01/42193 PCT/USOO/33057 --50preferably, phenyl or 4-hydroxy-3-hydroxymethylphenyl, more preferably phenyl. Yet another preferred group of compounds of Formula (I) are: 1. A compound of the formula: 5 OH H Arl

-

1 a1 H OH 10 wherein: Ar is selected from the group consisting of 4-amino-3,5-dichlorophenyl and 3-formylamino-4-hydroxyphenyl; and the stereochemistry at *C and **C is (RS) and (RS); (R) and (R); (S) and (S); (R) and (S); or (S) and (R); and pharmaceutically acceptable salts thereof. 15 Within this group, a more preferred compound is that wherein Arl is 4 amino-3,5-dichlorophenyl, and the stereochemistry at *C is (RS) and the stereochemistry at **C is (RS), or the stereochemistry at *C is (R) and the stereochemistry at **C is (R), or the stereochemistry at *C is (S) and the stereochemistry at **C is (S) or the stereochemistry at *C is (R) and the 20 stereochemistry at **C is (S) or the stereochemistry at *C is (S) and the stereochemistry at **C is (R). Within this group, another more preferred compound is that wherein Arl is 3-formylamino-4-hydroxyphenyl, and the stereochemistry at *C is (RS) and the stereochemistry at **C is (RS) or the stereochemistry at *C is (R) and the 25 stereochemistry at **C is (R) or the stereochemistry at *C is (S) and the stereochemistry at **C is (S) or the stereochemistry at *C is (R) and the stereochemistry at **C is (S) or the stereochemistry at *C is (S) and the stereochemistry at **C is (R). 30 WO 01/42193 PCT/USOO/33057 --5 1- GENERAL SYNTHETIC SCHEME Compounds of this invention can be made by the methods depicted in the reaction schemes shown below. The starting materials and reagents used in preparing these compounds are 5 either available from commercial suppliers such as Aldrich Chemical Co., (Milwaukee, Wisconsin, USA), Bachem (Torrance, California, USA), Emka-Chemie, or Sigma (St. Louis, Missouri, USA) or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-15 (John Wiley and 10 Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989), Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition), and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989). 15 The starting materials and the intermediates of the reaction may be isolated and purified if desired using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and the like. Such materials may be characterized using conventional means, including physical constants and spectral data. 20 Furthermore, it will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in 25 the art by routine optimization procedures. Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and 30 deprotection are well known in the art. For example, numerous protecting groups, WO 01/42193 PCT/USOO/33057 --52and their introduction and removal, are described in T. W. Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, and references cited therein. These schemes are merely illustrative of some methods by which the 5 compounds of this invention can be synthesized, and various modifications to these schemes can be made and will be suggested to one skilled in the art having referred to this disclosure. Preparation of a multibinding compound of Formula (I) 10 In general, a multibinding compound of Formula (I) where p is 2 and q is 1 can be prepared as illustrated and described in Schemes A-D below. A multibinding compound of Formula (I) where p is 2 and q is 1 can be prepared by covalently attaching the ligands, L, wherein at least one of the ligand is selected from a compound of formula (a) as defined in the Summary of the 15 Invention, to a linker, X, as shown in Scheme A below. Scheme A Method (a) 2 L FG+ FG 2

X-FG

2 . L yX L Method (b) LFG' + FG2- X -FG2PG

L

1 X -FG2PG [intermediate] (II) deprotect 2 FG' L1 X -- FG2 L? - L1 X -L2 20 In method (a), a multibinding compound of Formula (I) where p is 2 and q is 1 is prepared in one step, by covalently attaching the ligands, L, to a linker, X, where FG 1 and FG 2 represent a functional group such as halo, amino, hydroxy, thio, aldehyde, ketone, carboxy, carboxy derivatives such as acid halide, ester, WO 01/42193 PCT/USOO/33057 --53amido, and the like. This method is preferred for preparing compounds of Formula (I) where the ligands are the same. In method (b), the compounds of Formula (I) are prepared in a stepwise manner by covalently attaching one equivalent of a ligand, L,, with a ligand X 5 where where FG' and FG 2 represent a functional group as defined above, and

FG

2 PG is a protected functional group to give an intermediate of formula (II). Deprotection of the second functional group on the ligand, followed by reaction with a ligand L 2 , which may be same or different than ligand L,, then provides a compound of Formula (I). This method is suitable for preparing compounds of 10 Formula (I) where the ligands are the non-identical. The ligands are covalently attached to the linker using conventional chemical techniques providing for covalent linkage of the ligand to the linker. Reaction chemistries resulting in such linkages are well known in the art and involve the use of complementary functional groups on the linker and ligand as 15 shown in Table I below. Table I Representative Complementary Binding Chemistries First Reactive Group Second Reactive Group Linkage carboxyl amine amide 20 sulfonyl halide amine sulfonamide hydroxyl alkyl/aryl halide ether hydroxyl isocyanate urethane amine epoxide P-hydroxyamine amine alkyl/aryl halide alkylamine 25 amine isocyanate urea hydroxyl carboxyl ester amine aldehyde amine Reaction between a carboxylic acid of either the linker or the ligand and a primary or secondary amine of the ligand or the linker in the presence of suitable, 30 well-known activating agents such as dicyclohexylcarbodiimide, results in formation of an amide bond covalently linking the ligand to the linker; reaction between an amine group of either the linker or the ligand and a sulfonyl halide of WO 01/42193 PCT/USOO/33057 --54the ligand or the linker, in the presence of a base such as triethylamine, pyridine, an the like results in formation of a sulfonamide bond covalently linking the ligand to the linker; and reaction between an alcohol or phenol group of either the linker or the ligand and an alkyl or aryl halide of the ligand or the linker in the presence of a 5 base such as triethylamine, pyridine, and the like, results in formation of an ether bond covalently linking the ligand to the linker. A multibinding compound of Formula (I) where p is 2 q is 1, the second ligand Ar 3 is the same as Ar', X is a bond, and Q is 2-hydroxyethylamino group, 10 and the ligands are linked through the Ar2 group can be prepared from an aryl glyoxal derivative of formula 1 as shown in Scheme B below. Scheme B 0 0 0 2 Ar OH + H 2 N-W-Ar 2

-NH

2 : Ar1 N-W-Ar 2 --- N -s tKAr3 OH 1 2 3 I reduction OH H OH Ar 1 N-W-Ar 2 -HN Ar 3 (I) (Ar = Ar 3 ) Condensation of an acetophenone derivative of formula 1 with a diamine of formula 2 in an ethereal solution such as tetrahydrofuran provides an imine of 15 formula 3. Reduction of the imine with a suitable reducing agent such as diborane provides a compound of Formula (I). Suitable reaction solvents are tetrahydrofuran, and the like. Compound I where Ar' is phenyl is prepared by heating acetophenone in 48% hydrobromic acid in dimethylsulfoxide. Compounds of formula I can be prepared by methods well known in the art. 20 For example, ca-dihydroxy- 4-hydroxy-3-methoxycarbonylacetophenone can be prepared by heating 5--acetylsalicylic acid methyl ester in 48% hydrobromic acid.

WO 01/42193 PCT/USOO/33057 --55- Alternatively, a multibinding compound of Formula (I) where p is 2, q is 1, the second ligand Ar 3 is the same as Ar', X is a bond, and Q is 2 hydroxyethylamino group, and the ligands are linked through the Ar? group can be prepared from an aryl epoxide of formula 4 as shown in Scheme C below. 5 Scheme C 0 OH H_ OH + H 2 N-W-Ar 2

-NH

2 '-Ar N-W-Ar2-HN Ar3 4 2 (1) (Ar 1 = Ar 3 ) A compound of Formula (I) can be prepared by reacting an epoxide of formula 4 with a diamine of formula 2. Epoxides 4 are either commercially available or they can be prepared by the methods described in Kierstead, R.W. et. 10 al. J Med. Chem. 26, 1561-1569, (1983) or Hett, R. et. al. Tet. Lett.35, 9345-9348 (1994). Another method of preparing a multibinding compound of Formula (I) where p is 2, q is 1, the second ligand Ar3 is the same as Ar', X is a bond, and Q is 15 2-hydroxyethylamino group, and the ligands are linked through the A? group can be prepared from an acetophenone derivative of formula 5 as shown in Scheme D below. Scheme D WO 01/42193 PCT/USOO/33057 --56- Br 2 0 OH ArK Br2 'Ar1-KBr NaN3 , Ar!KN3 reduction Ar NH 2 CHCl 3 5 6 7 8 + OHC-W-Ar-CHO OH N-W-AHN Ar3 9 10 OH H OH Ar 1 NW-A-HN Ar 3 (I) (Ar 1 = Ar 3 ) Bromination of an acetophenone derivative of formula 5 with bromine in a halogenated organic solvent such as chloroform provides an a-bromoacetophenone derivative of formula 6. Treatment of 6 with sodium azide followed by reduction of 5 the resulting azide 7 with a suitable reducing agent such as lithium aluminum hydride provides ethanolamine derivative of formula 8. Condensation of two equivalents of 8 with a dialdehyde compound of formula 9 provides an imine of formula 10 which is converted to a compound of Formula (I) as described in Scheme A above. 10 Another method of preparing a multibinding compound of Formula (I) where p is 2, q is 1, Ar' and Ar 3 are different, X is a bond, and Q is 2 hydroxyethylamino group, and the ligands are linked through the A 2 group can be prepared as shown in Scheme E below. 15 Scheme E WO 01/42193 PCT/USOO/33057 --57- 0 0 Ar 3 OH + PG 2

PG

1 N-Ar 2

-W-HNPG

1 - PG 2

PG

1 N-W-Ar--N '-RAr3 OH 12 1 11 -2 OH PG 1 OH reductionOHPH r PG 2

PG

1 N-W-Ar 2

-NPG

1 Ar 3 -PG1HN-W-Ar 2 -N Ar 3 13 14 0 Ari1>KBr OH PG 1

PG

1 OH OH OH 6 Ar 1 N-W-Ar 2 _N Ar 3 Ar N-W-Ar 2 -NH Ar 3 ii reduction 15 (1) Condensation of a diamine of formula 11 (where PG, and PG 2 are suitable amino protecting groups which can be selectively removed) with a glyoxal of formula 2 followed by reduction of the resulting imine of formula 12 with a suitable reducing agent such as diborane in a suitable reaction solvents such as 5 tetrahydrofuran provides a compound of formula 13. Compounds of formula 11 can be prepared by methods described in figure 14. Reaction of compound 15 with an alpha bromoacetophenone compound of formula 6 followed by reduction of the keto group provides a compound of formula 16. The reaction is carried out under conditions well known in the art. 10 Deprotection of the amino protecting group then provides a compound of Formula (I). The deprotection reaction conditions depend on the nature of the protecting group. For example, if the protecting group is benzyl, it is removed under catalytic hydrogenation reaction conditions. 15 Any compound which is a P2 adrenergic receptor agonist can be used as a ligand in this invention. Typically, a compound selected for use as a ligand will have at least one functional group, such as an amino, hydroxyl, thiol or carboxyl group and the like, which allows the compound to be readily coupled to the linker. Compounds having such functionality are either known in the art or can be prepared WO 01/42193 PCT/USOO/33057 --58by routine modification of known compounds using conventional reagents and procedures. Linkers can be attached to different positions on the ligand molecule to achieve different orientations of the ligand domains, and thereby facilitate 5 multivalency. While a number of positions on p-adrenergic-modulating ligands are synthetically practical for linking, it is preferred to preserve those ligand substructures which are most important for ligand-receptor binding. At present, the aryl group and the sidechain nitrogen are preferred points of attachment. It will be apparent to one skilled in the art that the above chemistries are not 10 limited to preparing bivalent multibinding compounds of Formula (I) and can be used to prepare tri-, tetra-, etc., multibinding compounds of Formula (I). The linker is attached to the ligand at a position that retains ligand domain ligand binding site interaction and specifically which permits the ligand domain of the ligand to orient itself to bind to the ligand binding site. Such positions and 15 synthetic protocols for linkage are well known in the art. The term linker embraces everything that is not considered to be part of the ligand. The relative orientation in which the ligand domains are displayed derives from the particular point or points of attachment of the ligands to the linker, and on the framework geometry. The determination of where acceptable substitutions can 20 be made on a ligand is typically based on prior knowledge of structure-activity relationships (SAR) of the ligand and/or congeners and/or structural information about ligand-receptor complexes (e.g., X-ray crystallography, NMR, and the like). Such positions and the synthetic methods for covalent attachment are well known in the art. Following attachment to the selected linker (or attachment to a significant 25 portion of the linker, for example 2-10 atoms of the linker), the univalent linker ligand conjugate may be tested for retention of activity in the relevant assay. The linker, when covalently attached to multiple copies of the ligands, provides a biocompatible, substantially non-immunogenic multibinding compound. The biological activity of the multibinding compound is highly sensitive to the 30 valency, geometry, composition, size, flexibility or rigidity, etc. of the linker and, in turn, on the overall structure of the multibinding compound, as well as the presence or absence of anionic or cationic charge, the relative hydrophobicity/hydrophilicity WO 01/42193 PCT/USOO/33057 --59of the linker, and the like on the linker. Accordingly, the linker is preferably chosen to maximize the biological activity of the multibinding compound. The linker may be chosen to enhance the biological activity of the molecule. In general, the linker may be chosen from any organic molecule construct that orients two or more 5 ligands to their ligand binding sites to permit multivalency. In this regard, the linker can be considered as a "framework" on which the ligands are arranged in order to bring about the desired ligand-orienting result, and thus produce a multibinding compound. For example, different orientations can be achieved by including in the 10 framework groups containing mono- or polycyclic groups, including aryl and/or heteroaryl groups, or structures incorporating one or more carbon-carbon multiple bonds (alkenyl, alkenylene, alkynyl or alkynylene groups). Other groups can also include oligomers and polymers which are branched- or straight-chain species. In preferred embodiments, rigidity is imparted by the presence of cyclic groups (e.g., 15 aryl, heteroaryl, cycloalkyl, heterocyclic, etc.). In other preferred embodiments, the ring is a six or ten member ring. In still further preferred embodiments, the ring is an aromatic ring such as, for example, phenyl or naphthyl. Different hydrophobic/hydrophilic characteristics of the linker as well as the presence or absence of charged moieties can readily be controlled by the skilled 20 artisan. For example, the hydrophobic nature of a linker derived from hexamethylene diamine (H 2

N(CH

2

)

6 NH2) or related polyamines can be modified to be substantially more hydrophilic by replacing the alkylene group with a poly(oxyalkylene) group such as found in the commercially available "Jeffamines". Different frameworks can be designed to provide preferred orientations of 25 the ligands. Such frameworks may be represented by using an array of dots (as shown below) wherein each dot may potentially be an atom, such as C, 0, N, S, P, H, F, Cl, Br, and F or the dot may alternatively indicate the absence of an atom at that position. To facilitate the understanding of the framework structure, the framework is illustrated as a two dimensional array in the following diagram, 30 although clearly the framework is a three dimensional array in practice: WO 01/42193 PCT/USOO/33057 --60 8 - - - - - - - - -* -- 6 - - - - - - - - -* -- 5 - - - - - - - - -U - 4 - . . . . . - - -U - 3 - - - - - - - - -U - 2 - - - - * - 1 . - . . . . . - -U - 0 - - - . - - - - -* -- 0 12 3 4 5 6 7 8 Each dot is either an atom, chosen from carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus, or halogen, or the dot represents a point in space (i.e., an absence of an atom). As is apparent to the skilled artisan, only certain atoms on the 5 grid have the ability to act as an attachment point for the ligands, namely, C, 0, N, S and P. Atoms can be connected to each other via bonds (single, double or triple bonds with acceptable resonance and tautomeric forms), with regard to the usual constraints of chemical bonding. Ligands may be attached to the framework via 10 single, double or triple bonds (with chemically acceptable tautomeric and resonance forms). Multiple ligand groups (2 to 10) can be attached to the framework such that the minimal, shortest path distance between adjacent ligand groups does not exceed 100 atoms. Preferably, the linker connections to the ligand is selected such that the maximum spatial distance between two adjacent ligands is no more than 100A. 15 An example of a linker as presented by the grid is shown below for a biphenyl construct.

WO 01/42193 PCT/USOO/33057 --61- 8 - - - - - - - - - 6 -H H H 5 - - - . - - - - - 3 - - - -. 2 -H H H C 0 - - - - 0 1 2 3 4 5 6 7 8 9 10 Nodes (1,2), (2,0), (4,4), (5,2), (4,0), (6,2), (7,4), (9,4), (10,2), (9,0), (7,0) all represent carbon atoms. Node (10,0) represents a chlorine atom. All other nodes (or dots) are points in space (i.e., represent an absence of atoms). 5 Nodes (1,2) and (9,4) are attachment points. Hydrogen atoms are affixed to nodes (2,4), (4,4), (4,0), (2,0), (7,4), (10,2) and (7,0). Nodes (5,2) and (6,2) are connected by a single bond. The carbon atoms present are connected by either a single or double bonds, taking into consideration the principle of resonance and/or tautomerism. 10 The intersection of the framework (linker) and the ligand group, and indeed, the framework (linker) itself can have many different bonding patterns. Examples of acceptable patterns of three contiguous atom arrangements are shown in the following diagram: WO 01/42193 PCT/USOO/33057 --62- CCC NCC OCC SCC PCC CCN NCN OCN SCN PCN CCO NO OCO SC0 PCO CCS NCS OCS SCS PCS CCP NCP OCP SCP PC P CNC NNC ONC SNC P NC CNN NNN ONN SNN PNN CNO NNO ONO S9-R- PNO CNS NNS DT~S~ SNS PNS CNP NTP ONP SNP PNP C0C N OO C S 0C POC coo NON DDR SON PON COC= 0-0 000 Soo POO C O0 P PTZ 0= D-~ - 5 -0-S 050 ND O P -0-P p---P C SC N SC T- -T CSN NSN OSC SSC PSC CSO NSO OSN SSN PSN CSS NSS OSO SSO p-9U CSP NSP OSS 5-55 p-S-g OSP 5S-~P p-s-P CPC NPC CPN NPN OPC SPC PPC CPO NPO OPN SPN p-P-R CPS NPS OPO SPO p-pD CPP NPP OPS SPS p-p-S OPP SPP p-p One skilled in the art would be able to identify bonding patterns that would produce multivalent compounds. Methods for producing these bonding arrangements are described in March, "Advanced Organic Chemistry", 4th Edition, Wiley-Interscience, New York, New York (1992). These arrangements are 5 described in the grid of dots shown in the scheme above. All of the possible arrangements for the five most preferred atoms are shown. Each atom has a variety of acceptable oxidation states. The bonding arrangements underlined are less acceptable and are not preferred. Examples of molecular structures in which the above bonding patterns could 10 be employed as components of the linker are shown below. 15 WO 01/42193 PCT/USOO/33057 --63- N N' N C kCC' 0 10 0 0 0 C C C S N.'S N S N' C C 0 NO.~ O O\ O 0 NO O N C N C C 15 0 NN N N CC, N, 0CN NN N 0 0 NC C' N CC NN T 0 00 N- N ONON h 20 e identification of an appropriate framework geometry and size for ligand domain presentation are important steps in the construction of a multibinding compound with enhanced activity. Systematic spatial searching strategies can be used to aid in the identification of preferred frameworks through an iterative process. Figure 3 illustrates a useful strategy for determining an optimal framework display 25 orientation for ligand domains. Various other strategies are known to those skilled in the art of molecular design and can be used for preparing compounds of this invention. As shown in Figure 1, display vectors around similar central core structures such as a phenyl structure (Panel A) and a cyclohexane structure (Panel B) can be 30 varied, as can the spacing of the ligand domain from the core structure (i.e., the length of the attaching moiety). It is to be noted that core structures other than those shown here can be used for determining the optimal framework display WO 01/42193 PCT/USOO/33057 --64orientation of the ligands. The process may require the use of multiple copies of the same central core structure or combinations of different types of display cores. The above-described process can be extended to trimers (Figure 2) and compound of higher valency (Figures 3 and 4). 5 Assays of each of the individual compounds of a collection generated as described above will lead to a subset of compounds with the desired enhanced activities (e.g., potency, selectivity, etc.). The analysis of this subset using a technique such as Ensemble Molecular Dynamics will provide a framework orientation that favors the properties desired. A wide diversity of linkers is 10 commercially available (see, e.g., Available Chemical Directory (ACD)). Many of the linkers that are suitable for use in this invention fall into this category. Other can be readily synthesized by methods well known in the art and/or are described below. Having selected a preferred framework geometry, the physical properties of 15 the linker can be optimized by varying the chemical composition thereof. The composition of the linker can be varied in numerous ways to achieve the desired physical properties for the multibinding compound. It can therefore be seen that there is a plethora of possibilities for the composition of a linker. Examples of linkers include aliphatic moieties, aromatic 20 moieties, steroidal moieties, peptides, and the like. Specific examples are peptides or polyamides, hydrocarbons, aromatic groups, ethers, lipids, cationic or anionic groups, or a combination thereof. Examples are given below, but it should be understood that various changes may be made and equivalents may be substituted without departing from the true 25 spirit and scope of the invention. For example, properties of the linker can be modified by the addition or insertion of ancillary groups into or onto the linker, for example, to change the solubility of the multibinding compound (in water, fats, lipids, biological fluids, etc.), hydrophobicity, hydrophilicity, linker flexibility, antigenicity, stability, and the like. For example, the introduction of one or more 30 poly(ethylene glycol) (PEG) groups onto or into the linker enhances the hydrophilicity and water solubility of the multibinding compound, increases both molecular weight and molecular size and, depending on the nature of the WO 01/42193 PCT/USOO/33057 --65unPEGylated linker, may increase the in vivo retention time. Further PEG may decrease antigenicity and potentially enhances the overall rigidity of the linker. Ancillary groups which enhance the water solubility/hydrophilicity of the linker and, accordingly, the resulting multibinding compounds are useful in 5 practicing this invention. Thus, it is within the scope of the present invention to use ancillary groups such as, for example, small repeating units of ethylene glycols, alcohols, polyols (e.g., glycerin, glycerol propoxylate, saccharides, including mono , oligosaccharides, etc.), carboxylates (e.g., small repeating units of glutamic acid, acrylic acid, etc.), amines (e.g., tetraethylenepentamine), and the like) to enhance 10 the water solubility and/or hydrophilicity of the multibinding compounds of this invention. In preferred embodiments, the ancillary group used to improve water solubility/hydrophilicity will be a polyether. The incorporation of lipophilic ancillary groups within the structure of the linker to enhance the lipophilicity and/or hydrophobicity of the multibinding 15 compounds described herein is also within the scope of this invention. Lipophilic groups useful with the linkers of this invention include, by way of example only, aryl and heteroaryl groups which, as above, may be either unsubstituted or substituted with other groups, but are at least substituted with a group which allows their covalent attachment to the linker. Other lipophilic groups useful with the 20 linkers of this invention include fatty acid derivatives which do not form bilayers in aqueous medium until higher concentrations are reached. Also within the scope of this invention is the use of ancillary groups which result in the multibinding compound being incorporated or anchored into a vesicle or other membranous structure such as a liposome or a micelle. The term "lipid" 25 refers to any fatty acid derivative that is capable of forming a bilayer or a micelle such that a hydrophobic portion of the lipid material orients toward the bilayer while a hydrophilic portion orients toward the aqueous phase. Hydrophilic characteristics derive from the presence of phosphato, carboxylic, sulfato, amino, sulfhydryl, nitro and other like groups well known in the art. Hydrophobicity could 30 be conferred by the inclusion of groups that include, but are not limited to, long chain saturated and unsaturated aliphatic hydrocarbon groups of up to 20 carbon atoms and such groups substituted by one or more aryl, heteroaryl, cycloalkyl, WO 01/42193 PCT/USOO/33057 --66and/or heterocyclic group(s). Preferred lipids are phosphglycerides and sphingolipids, representative examples of which include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyleoyl phosphatidylcholine, lysophosphatidylcholine, 5 lysophosphatidylethanolamine, dipalmitoylphosphatidylcholine, dioleoyl phosphatidylcholine, distearoyl-phosphatidylcholine or dilinoleoylphosphatidyl choline could be used. Other compounds lacking phosphorus, such as sphingolipid and glycosphingolipid families are also within the group designated as lipid. Additionally, the amphipathic lipids described above may be mixed with other 10 lipids including triglycerides and sterols. The flexibility of the linker can be manipulated by the inclusion of ancillary groups which are bulky and/or rigid. The presence of bulky or rigid groups can hinder free rotation about bonds in the linker or bonds between the linker and the ancillary group(s) or bonds between the linker and the functional groups. Rigid 15 groups can include, for example, those groups whose conformational lability is restrained by the presence of rings and/or multiple bonds within the group, for example, aryl, heteroaryl, cycloalkyl, cycloalkenyl, and heterocyclic groups. Other groups which can impart rigidity include polypeptide groups such as oligo- or polyproline chains. 20 Rigidity can also be imparted electrostatically. Thus, if the ancillary groups are either positively or negatively charged, the similarly charged ancillary groups will force the presenter linker into a configuration affording the maximum distance between each of the like charges. The energetic cost of bringing the like-charged groups closer to each other will tend to hold the linker in a configuration that 25 maintains the separation between the like-charged ancillary groups. Further ancillary groups bearing opposite charges will tend to be attracted to their oppositely charged counterparts and potentially may enter into both inter- and intramolecular ionic bonds. This non-covalent mechanism will tend to hold the linker into a conformation which allows bonding between the oppositely charged 30 groups. The addition of ancillary groups which are charged, or alternatively, bear a latent charge when deprotected, following addition to the linker, include deprotonation of a carboxyl, hydroxyl, thiol or amino group by a change in pH, WO 01/42193 PCT/USOO/33057 --67oxidation, reduction or other mechanisms known to those skilled in the art which result in removal of the protecting group, is within the scope of this invention. Rigidity may also be imparted by internal hydrogen bonding or by hydrophobic collapse. 5 Bulky groups can include, for example, large atoms, ions (e.g., iodine, sulfur, metal ions, etc.) or groups containing large atoms, polycyclic groups, including aromatic groups, non-aromatic groups and structures incorporating one or more carbon-carbon multiple bonds (i.e., alkenes and alkynes). Bulky groups can also include oligomers and polymers which are branched- or straight-chain species. 10 Species that are branched are expected to increase the rigidity of the structure more per unit molecular weight gain than are straight-chain species. In preferred embodiments, rigidity is imparted by the presence of cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, heterocyclic, etc.). In other preferred embodiments, the linker comprises one or more six-membered rings. In still further 15 preferred embodiments, the ring is an aryl group such as, for example, phenyl or naphthyl. In view of the above, it is apparent that the appropriate selection of a linker group providing suitable orientation, restricted/unrestricted rotation, the desired degree of hydrophobicity/hydrophilicity, etc. is well within the skill of the art. 20 Eliminating or reducing antigenicity of the multibinding compounds described herein is also within the scope of this invention. In certain cases, the antigenicity of a multibinding compound may be eliminated or reduced by use of groups such as, for example, poly(ethylene glycol). As explained above, the multibinding compounds described herein comprise 25 2-10 ligands attached to a linker that attaches the ligands in such a manner that they are presented to the enzyme for multivalent interactions with ligand binding sites thereon/therein. The linker spatially constrains these interactions to occur within dimensions defined by the linker. This and other factors increases the biological activity of the multibinding compound as compared to the same number of ligands 30 made available in monobinding form. The compounds of this invention are preferably represented by the empirical Formula (L)P(X)q where L, X, p and q are as defined above. This is intended to WO 01/42193 PCT/USOO/33057 --68include the several ways in which the ligands can be linked together in order to achieve the objective of multivalency, and a more detailed explanation is described below. As noted previously, the linker may be considered as a framework to which 5 ligands are attached. Thus, it should be recognized that the ligands can be attached at any suitable position on this framework, for example, at the termini of a linear chain or at any intermediate position. The simplest and most preferred multibinding compound is a bivalent compound which can be represented as L-X-L, where each L is independently a 10 ligand which may be the same or different and each X is independently the linker. Examples of such bivalent compounds are provided in FIG. 1 where each shaded circle represents a ligand. A trivalent compound could also be represented in a linear fashion, i.e., as a sequence of repeated units L-X-L-X-L, in which L is a ligand and is the same or different at each occurrence, as can X. However, a trimer 15 can also be a radial multibinding compound comprising three ligands attached to a central core, and thus represented as (L) 3 X, where the linker X could include, for example, an aryl or cycloalkyl group. Illustrations of trivalent and tetravalent compounds of this invention are found in FIG.s 2 and 3 respectively where, again, the shaded circles represent ligands. Tetravalent compounds can be represented in a 20 linear array, e.g., L-X-L-X-L-X-L in a branched array, e.g., 25 L-X-L-X-L L 30 (a branched construct analogous to the isomers of butane -- n-butyl, iso-butyl, sec butyl, and t-butyl) or in a tetrahedral array, e.g., WO 01/42193 PCT/USOO/33057 --69- L L x L L where X and L are as defined herein. Alternatively, it could be represented as an alkyl, aryl or cycloalkyl derivative as above with four (4) ligands attached to the core linker. The same considerations apply to higher multibinding compounds of this 5 invention containing 5-10 ligands as illustrated in FIG. 4 where, as before, the shaded circles represent ligands. However, for multibinding agents attached to a central linker such as aryl or cycloalkyl, there is a self-evident constraint that there must be sufficient attachment sites on the linker to accommodate the number of ligands present; for example, a benzene ring could not directly accommodate more 10 than 6 ligands, whereas a multi-ring linker (e.g., biphenyl) could accommodate a larger number of ligands. The above described compounds may alternatively be represented as cyclic chains of the form: rL> x x KL and variants thereof. 15 All of the above variations are intended to be within the scope of the invention defined by the Formula (L)p(X)q. With the foregoing in mind, a preferred linker may be represented by the following formula: -Xa-Z-(Ya-Z)m-Xa_ 20 wherein m is an integer of from 0 to 20; Xa at each separate occurrence is selected from the group consisting of WO 01/42193 PCT/USOO/33057 --70- -0-, -S-, -NR-, -C(O)-, -C(O)0-, -OC(O)-, -C(O)NR-, -NRC(O)-, C(S), -C(S)O-, -C(S)NR-, -NRC(S)-, or a covalent bond where R is as defined below; Z at each separate occurrence is selected from the group consisting of alkylene, substituted alkylene, cycloalkylene, substituted cylcoalkylene, alkenylene, 5 substituted alkenylene, alkynylene, substituted alkynylene, cycloalkenylene, substituted cycloalkenylene, arylene, heteroarylene, heterocyclene, or a covalent bond; each ya at each separate occurrence is selected from the group consisting of -0-, -C(O)-, -OC(O)-, -C(0)O-, -NR-, -S(O)n-, -C(O)NR'-, -NR'C(O)-, 10 -NR'C(O)NR'-, -NR'C(S)NR'-, -C(=NR')-NR'-, -NR'-C(=NR')-, -OC(O)-NR'-, -NR'-C(O)-O-, -N=C(Xa)-NR'-, -NR'-C(Xa)=N-,-P(O)(OR')-O-, -O-P(O)(OR')-, -S(O).CR' R"-, -S(O)n-NR'-, -NR'-S(O), -, -S-S-, and a covalent bond; where n is 0, 1 or 2; and R, R' and R" at each separate occurrence are selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, 15 alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl and heterocyclic. Additionally, the linker moiety can be optionally substituted at any atom therein by one or more alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, 20 substituted alkynyl, aryl, heteroaryl and heterocyclic group. In view of the above description of the linker, it is understood that the term "linker" when used in combination with the term "multibinding compound" includes both a covalently contiguous single linker (e.g., L-X-L) and multiple covalently non-contiguous linkers (L-X-L-X-L) within the multibinding compound. 25 Combinatorial Libraries The methods described above lend themselves to combinatorial approaches for identifying multimeric compounds which possess multibinding properties. Specifically, factors such as the proper juxtaposition of the individual 30 ligands of a multibinding compound with respect to the relevant array of binding sites on a target or targets is important in optimizing the interaction of the multibinding compound with its target(s) and to maximize the biological advantage WO 01/42193 PCT/USOO/33057 --71 through multivalency. One approach is to identify a library of candidate multibinding compounds with properties spanning the multibinding parameters that are relevant for a particular target. These parameters include: (1) the identity of ligand(s), (2) the orientation of ligands, (3) the valency of the construct, (4) linker 5 length, (5) linker geometry, (6) linker physical properties, and (7) linker chemical functional groups. Libraries of multimeric compounds potentially possessing multibinding properties (i.e., candidate multibinding compounds) and comprising a multiplicity of such variables are prepared and these libraries are then evaluated via 10 conventional assays corresponding to the ligand selected and the multibinding parameters desired. Considerations relevant to each of these variables are set forth below: Selection of ligand(s): A single ligand or set of ligands is (are) selected for incorporation into the 15 libraries of candidate multibinding compounds which library is directed against a particular biological target or targets e.g., P2 adrenergic receptor. The only requirement for the ligands chosen is that they are capable of interacting with the selected target(s). Thus, ligands may be known drugs, modified forms of known drugs, substructures of known drugs or substrates of modified forms of known 20 drugs (which are competent to interact with the target), or other compounds. Ligands are preferably chosen based on known favorable properties that may be projected to be carried over to or amplified in multibinding forms. Favorable properties include demonstrated safety and efficacy in human patients, appropriate PK/ADME profiles, synthetic accessibility, and desirable physical properties such 25 as solubility, log P, etc. However, it is crucial to note that ligands which display an unfavorable property from among the previous list may obtain a more favorable property through the process of multibinding compound formation; i.e., ligands should not necessarily be excluded on such a basis. For example, a ligand that is not sufficiently potent at a particular target so as to be efficacious in a human 30 patient may become highly potent and efficacious when presented in multibinding form. A ligand that is potent and efficacious but not of utility because of a non mechanism-related toxic side effect may have increased therapeutic index WO 01/42193 PCT/USOO/33057 --72- (increased potency relative to toxicity) as a multibinding compound. Compounds that exhibit short in vivo half-lives may have extended half-lives as multibinding compounds. Physical properties of ligands that limit their usefulness (e.g. poor bioavailability due to low solubility, hydrophobicity, hydrophilicity) may be 5 rationally modulated in multibinding forms, providing compounds with physical properties consistent with the desired utility. Orientation: selection of ligand attachment points and linking chemistry: Several points are chosen on each ligand at which to attach the ligand to the linker. The selected points on the ligand/linker for attachment are functionalized to 10 contain complementary reactive functional groups. This permits probing the effects of presenting the ligands to their receptor(s) in multiple relative orientations, an important multibinding design parameter. The only requirement for choosing attachment points is that attaching to at least one of these points does not abrogate activity of the ligand. Such points for attachment can be identified by structural 15 information when available. For example, inspection of a co-crystal structure of a protease inhibitor bound to its target allows one to identify one or more sites where linker attachment will not preclude the enzyme:inhibitor interaction. Alternatively, evaluation of ligand/target binding by nuclear magnetic resonance will permit the identification of sites non-essential for ligand/target binding. See, for example, 20 Fesik, et al., U.S. Patent No. 5,891,643. When such structural information is not available, utilization of structure-activity relationships (SAR) for ligands will suggest positions where substantial structural variations are and are not allowed. In the absence of both structural and SAR information, a library is merely selected with multiple points of attachment to allow presentation of the ligand in multiple 25 distinct orientations. Subsequent evaluation of this library will indicate what positions are suitable for attachment. It is important to emphasize that positions of attachment that do abrogate the activity of the monomeric ligand may also be advantageously included in candidate multibinding compounds in the library provided that such compounds bear at least 30 one ligand attached in a manner which does not abrogate intrinsic activity. This selection derives from, for example, heterobivalent interactions within the context of a single target molecule. For example, consider a receptor antagonist ligand WO 01/42193 PCT/USOO/33057 --73bound to its target receptor, and then consider modifying this ligand by attaching to it a second copy of the same ligand with a linker which allows the second ligand to interact with the same receptor molecule at sites proximal to the antagonist binding site, which include elements of the receptor that are not part of the formal 5 antagonist binding site and/or elements of the matrix surrounding the receptor such as the membrane. Here, the most favorable orientation for interaction of the second ligand molecule with the receptor/matrix may be achieved by attaching it to the linker at a position which abrogates activity of the ligand at the formal antagonist binding site. Another way to consider this is that the SAR of individual 10 ligands within the context of a multibinding structure is often different from the SAR of those same ligands in momomeric form. The foregoing discussion focused on bivalent interactions of dimeric compounds bearing two copies of the same ligand joined to a single linker through different attachment points, one of which may abrogate the binding/activity of the 15 monomeric ligand. It should also be understood that bivalent advantage may also be attained with heterodimeric constructs bearing two different ligands that bind to common or different targets. For example, a 5HT 4 receptor antagonist and a bladder-selective muscarinic M 3 antagonist may be joined to a linker through attachment points which do not abrogate the binding affinity of the monomeric 20 ligands for their respective receptor sites. The dimeric compound may achieve enhanced affinity for both receptors due to favorable interactions between the 5HT 4 ligand and elements of the M 3 receptor proximal to the formal M 3 antagonist binding site and between the M 3 ligand and elements of the 5HT 4 receptor proximal to the formal 5HT 4 antagonist binding site. Thus, the dimeric compound may be 25 more potent and selective antagonist of overactive bladder and a superior therapy for urinary urge incontinence. Once the ligand attachment points have been chosen, one identifies the types of chemical linkages that are possible at those points. The most preferred types of chemical linkages are those that are compatible with the overall structure of the 30 ligand (or protected forms of the ligand) readily and generally formed, stable and intrinsically inocuous under typical chemical and physiological conditions, and WO 01/42193 PCT/USOO/33057 --74compatible with a large number of available linkers. Amide bonds, ethers, amines, carbamates, ureas, and sulfonamides are but a few examples of preferred linkages. Linkers: spanning relevant multibinding parameters through selection of valency, linker length, linker geometry, rigidity, physical properties, and chemical functional 5 groups In the library of linkers employed to generate the library of candidate multibinding compounds, the selection of linkers employed in this library of linkers takes into consideration the following factors: Valency: 10 In most instances the library of linkers is initiated with divalent linkers. The choice of ligands and proper juxtaposition of two ligands relative to their binding sites permits such molecules to exhibit target binding affinities and specificities more than sufficient to confer biological advantage. Furthermore, divalent linkers or constructs are also typically of modest size such that they retain the desirable 15 biodistribution properties of small molecules. Linker length: Linkers are chosen in a range of lengths to allow the spanning of a range of inter-ligand distances that encompass the distance preferable for a given divalent interaction. In some instances the preferred distance can be estimated rather 20 precisely from high-resolution structural information of targets, typically enzymes and soluble receptor targets. In other instances where high-resolution structural information is not available (such as 7TM G-protein coupled receptors), one can make use of simple models to estimate the maximum distance between binding sites either on adjacent receptors or at different locations on the same receptor. In 25 situations where two binding sites are present on the same target (or target subunit for multisubunit targets), preferred linker distances are 2-20 A, with more preferred linker distances of 3-12 A. In situations where two binding sites reside on separate (e.g., protein) target sites, preferred linker distances are 20-100 A, with more preferred distances of 30-70 A. 30 Linker geometry and rigidity: The combination of ligand attachment site, linker length, linker geometry, and linker rigidity determine the possible ways in which the ligands of candidate WO 01/42193 PCT/USOO/33057 --75multibinding compounds may be displayed in three dimensions and thereby presented to their binding sites. Linker geometry and rigidity are nominally determined by chemical composition and bonding pattern, which may be controlled and are systematically varied as another spanning function in a multibinding array. 5 For example, linker geometry is varied by attaching two ligands to the ortho, meta, and para positions of a benzene ring, or in cis- or trans-arrangements at the 1,1 - vs. 1,2- vs. 1,3- vs. 1,4- positions around a cyclohexane core or in cis- or trans arrangements at a point of ethylene unsaturation. Linker rigidity is varied by controlling the number and relative energies of different conformational states 10 possible for the linker. For example, a divalent compound bearing two ligands joined by 1,8-octyl linker has many more degrees of freedom, and is therefore less rigid than a compound in which the two ligands are attached to the 4,4' positions of a biphenyl linker. Linker physical properties: 15 The physical properties of linkers are nominally determined by the chemical constitution and bonding patterns of the linker, and linker physical properties impact the overall physical properties of the candidate multibinding compounds in which they are included. A range of linker compositions is typically selected to provide a range of physical properties (hydrophobicity, hydrophilicity, 20 amphiphilicity, polarization, acidity, and basicity) in the candidate multibinding compounds. The particular choice of linker physical properties is made within the context of the physical properties of the ligands they join and preferably the goal is to generate molecules with favorable PK/ADME properties. For example, linkers can be selected to avoid those that are too hydrophilic or too hydrophobic to be 25 readily absorbed and/or distributed in vivo. Linker chemical functional groups: Linker chemical functional groups are selected to be compatible with the chemistry chosen to connect linkers to the ligands and to impart the range of physical properties sufficient to span initial examination of this parameter. 30 Combinatorial synthesis WO 01/42193 PCT/USOO/33057 --76- Having chosen a set of n ligands (n being determined by the sum of the number of different attachment points for each ligand chosen) and m linkers by the process outlined above, a library of (n!)m candidate divalent multibinding compounds is prepared which spans the relevant multibinding design parameters for 5 a particular target. For example, an array generated from two ligands, one which has two attachment points (A1, A2) and one which has three attachment points (B 1, B2, B3) joined in all possible combinations provide for at least 15 possible combinations of multibinding compounds: 10 Al-Al Al-A2 Al-Bl Al-B2 Al-B3 A2-A2 A2-B1 A2-B2 A2-B3 B1-Bl B1-B2 Bl-B3 B2-B2 B2-B3 B3-B3 When each of these combinations is joined by 10 different linkers, a library of 150 candidate multibinding compounds results. 15 Given the combinatorial nature of the library, common chemistries are preferably used to join the reactive functionalies on the ligands with complementary reactive functionalities on the linkers. The library therefore lends itself to efficient parallel synthetic methods. The combinatorial library can employ solid phase chemistries well known in the art wherein the ligand and/or linker is attached to a 20 solid support. Alternatively and preferably, the combinatorial libary is prepared in the solution phase. After synthesis, candidate multibinding compounds are optionally purified before assaying for activity by, for example, chromatographic methods (e.g., HPLC). Analysis of array by biochemical, analytical, pharmacological, and computational 25 methods: Various methods are used to characterize the properties and activities of the candidate multibinding compounds in the library to determine which compounds possess multibinding properties. Physical constants such as solubility under various solvent conditions and logD/clogD values can be determined. A combination of 30 NMR spectroscopy and computational methods is used to determine low-energy conformations of the candidate multibinding compounds in fluid media. The ability of the members of the library to bind to the desired target and other targets is WO 01/42193 PCT/USOO/33057 --77determined by various standard methods, which include radioligand displacement assays for receptor and ion channel targets, and kinetic inhibition analysis for many enzyme targets. In vitro efficacy, such as for receptor agonists and antagonists, ion channel blockers, and antimicrobial activity, can also be determined. 5 Pharmacological data, including oral absorption, everted gut penetration, other pharmacokinetic parameters and efficacy data can be determined in appropriate models. In this way, key structure-activity relationships are obtained for multibinding design parameters which are then used to direct future work. The members of the library which exhibit multibinding properties, as 10 defined herein, can be readily determined by conventional methods. First those members which exhibit multibinding properties are identified by conventional methods as described above including conventional assays (both in vitro and in vivo). Second, ascertaining the structure of those compounds which exhibit 15 multibinding properties can be accomplished via art recognized procedures. For example, each member of the library can be encrypted or tagged with appropriate information allowing determination of the structure of relevant members at a later time. See, for example, Dower, et al., International Patent Application Publication No. WO 93/06121; Brenner, et al., Proc. Natl. Acad. Sci., USA, 89:5181 (1992); 20 Gallop, et al., U.S. Patent No. 5,846,839; each of which are incorporated herein by reference in its entirety. Alternatively, the structure of relevant multivalent compounds can also be determined from soluble and untagged libaries of candidate multivalent compounds by methods known in the art such as those described by Hindsgaul, et al., Canadian Patent Application No. 2,240,325 which was published 25 on July 11, 1998. Such methods couple frontal affinity chromatography with mass spectroscopy to determine both the structure and relative binding affinities of candidate multibinding compounds to receptors. The process set forth above for dimeric candidate multibinding compounds can, of course, be extended to trimeric candidate compounds and higher analogs 30 thereof. Follow-up synthesis and analysis of additional array(s): WO 01/42193 PCT/USOO/33057 --78- Based on the information obtained through analysis of the initial library, an optional component of the process is to ascertain one or more promising multibinding "lead" compounds as defined by particular relative ligand orientations, linker lengths, linker geometries, etc. Additional libraries can then be generated 5 around these leads to provide for further information regarding structure to activity relationships. These arrays typically bear more focused variations in linker structure in an effort to further optimize target affinity and/or activity at the target (antagonism, partial agonism, etc.), and/or alter physical properties. By iterative redesign/analysis using the novel principles of multibinding design along with 10 classical medicinal chemistry, biochemistry, and pharmacology approaches, one is able to prepare and identify optimal multibinding compounds that exhibit biological advantage towards their targets and as therapeutic agents. To further elaborate upon this procedure, suitable divalent linkers include, by way of example only, those derived from dicarboxylic acids, disulfonylhalides, 15 dialdehydes, diketones, dihalides, diisocyanates,diamines, diols, mixtures of carboxylic acids, sulfonylhalides, aldehydes, ketones, halides, isocyanates, amines and diols. In each case, the carboxylic acid, sulfonylhalide, aldehyde, ketone, halide, isocyanate, amine and diol functional group is reacted with a complementary functionality on the ligand to form a covalent linkage. Such complementary 20 functionality is well known in the art as illustrated in the following table: COMPLEMENTARY BINDING CHEMISTRIES First Reactive Group Second Reactive Group Linkage 25 hydroxyl isocyanate urethane amine epoxide P-hydroxyamine hydroxyamine sulfonyl halide sulfonamide carboxyl acid amine amide hydroxyl alkyl/aryl halide ether 30 aldehyde amine/NaCNBH 3 amine ketone amine/NaCNBH 3 amine amine isocyanate urea WO 01/42193 PCT/USOO/33057 --79- The following table illustrates, by way of examples, starting materials (identified as X- 1 through X-418) that can be used to prepare linkers incorporated in the multibinding compounds of this invention utilizing the chemistry described above. For example, 1,10-decanedicarboxylic acid, X1, can be reacted with 2 5 equivalents of a ligand carrying an amino group in the presence of a coupling reagent such as DCC to provide a bivalent multibinding compound of formula (I) wherein the ligands are linked via 1,1 0-decanediamido linking group.

WO 01/42193 PCT/USOO/33057 --80- Diacids 0 OH HO H HO 0 X-2 0 OHOH 0. _0 0OH HO 0 HO0 x3OH HjC C/-I 3 O'' X-3.X CH3 0 OH CH X4 X-5 OLO OH HD 0 0 O 0O HOH0 X-6 HOX-7 HO 0 - 8 0 OH OH OH HO 0 OH \OH N 0 HO 0 HO N HO CHJ HO CH 3 H2 0 X-9 X-10 X-11 OH HO 0 HO 0 OH X-14 X-13 HO OH HO HO lI - X-15 Xo6X1 OHOH

H

3 C H

H

3 C X-18 X1 OH X- 19 WO 01/42193 PCT/USOO/33057 --81- Diacids 0 OH H O

CH

3 OHO HO 0 X-2 0 OH OH 0 0 0 0 OH OH HJC O) O OH

CH

3 X- 4 CjX-5 L/o S('~O 0 OH

.

oo OH X-6 HO X-7 HO, 0 0 X-8 - OH OH OH HO 0 OH OH N O HO O HO HO HO CHj HO CH 3 X-12 0 X-9 X-10 X-11 O OH 0 H0 00 OH X-14 X-13 0 OHOH 0 OHO HO HHO OH - HfC OH 3 X-15 X- 16 0 X-17 OHOH 00

H

3 C 0

H

3 C X-18 OH. X- 19 WO 01/42193 PCT/USOO/33057 82- OH H HO 0 OH HO. HO 0-, 0 X-20 X-21 HO H X-22 0 0 0 HO OH 0 OH 0= .OH X-24 0 X-23 0 HO X-5OH X-25 OH OHO X-26 00 Chiral N OH X-27 o 0 HO 00 H OH 0 H HO OH H CH 2 0 X-29 OH X-28 X-JO HOH HOH OH o 0 X-31 HO N O 0 N OH 0 Chiral X-32 WO 01/42193 PCT/USOO/33057 --83- 0 0 0 OH 0 0 H OX HO X-363 00 HO,- OH CH NOH 0 0 0 HO0 HOF F'X-37 JO X-8 H X-36 0 OH C lChira X-40 X-41 H-3C NOH 3 00 HOH HO HO0 HoI~i~ HO F4 3 C7 HOH X-42 OH4 0 OH HO 0 o 0 HOO CH O OH OH OH O X-45 X-46 Chiral H 3 C OH ' X-47 X-48 WO 01/42193 PCT/USOO/33057 --84- HO 0 O Cl-y OH. X-49 HO 0 HO OH OH 0 0 N~H 0=>F0Cia O N S-S N n H HO X-51 X-52 Chiral F F X-50

H

2 N HN 0 N N HN 00 H 0 OH 0 ~~ 0 OH OH OH HX-55 HO Chirol HO Chiral X-53 X-54 0 0 0 OH

CH

3 OH HO C -X-57 X-58

H

3 C X-56 O 0 OH HO 0 HO 0 O O Chiral XC60 X-59X 0 ~ OHO 0 0 0 H HOH

HO

H Shir0 Chiral X-61 X-62 WO 01/42193 PCT/USOO/33057 --85- 00 HO C C O(

H

3 C 00 NH 0OH >- OH HCOj

H

3 C OH HO S S HO HO 0 Chiral 6 X-66 X-64 HO OH OH HO HO ~ HO0 X80NX-67 0 X66 OH HO 0 0 0 0 0F N. OH 0.0 HO HOOH 0 \/-0 CN Chirol HOO X-69 X 70 HO HHFFFFFFF 0 Hir HO HOOFHFFO X-7 7 X-72 X-7 HO HO 0 OH HO 0 0 HO ~S 0 0 X-75 X7 X-74 00 OH O 0 I-1 3 C HO. HO X- 77 X- 78.

WO 01/42193 PCT/USOO/33057 --86- C 0 N ChiraOH Chira CH 0-80 X-81 ~ O 90 0 0 OH OH H0 N OH HOOH HOH 'Chiral X-83 X-84 X-82 WO 01/42193 PCT/USOO/33057 -- 87 00 X-85 OH CH3 00 N N OH H 00 HO <~I...,OH H OH HO~0Chiral 0X-88 HO OX-8 OH-8 HOOHoOH 0 NHHO N N H - 0 X-89 X-90 0 00 OH OH HO H3C O

CH

3 OH Chiral X-92 0 0 0 FF FFFF FFFF O N 0 OH HH HO OH HOFF FE FF FE FEF OH X6X X-9 X-96 WO 01/42193 PCT/USOO/33057 --88- HO 0 OH FF0 0 0 HJ CCHj HJCCHJ + 3 N OH HO OH HXN 0 0 HO X-98 X-99 X-X100 HQ 0 FF.FF FFFF OH O NHO 00 H O OH HOX-1Ol 0 0 FFFFFF 0 X-X102 0. N H0 OH 0 (NyOH 0 OH N OH O HO ~ 0 ss, 0 OH 0 X-105 X-104 X0 0 0 N 0 CN OH COH 0 OH OH N 00:) HO 0 X- 108 X0 a c/ X- 107C WO 01/42193 PCT/USOO/33057 -89- 0 0 OH N OH SHO N OH 0 HOo C/, OH CHj j Chiral Br X-111 Chiral X-110 X-112 0 OH OH 00 HO OH 0 0 0HO 00 OH OHO H H 0 HO OH HO OH Chiral X -115 0 0 " Z HO HO N H 0 N O OH OH 00 X-117 X-116 0 0 OH OH HO0 O O OH o X-118 0 X- 120 X-119 0 X-122 00 SS. 0 OHO 0 N H X-124 0 ~ Chiol 0H 2 N X-125 X-126 X-123 WO 01/42193 PCT/USOO/33057 --90- OH HOH ~OH Ho HO X-128 0 N OH OH y 0 HO'f' 'O ,,_O 0 C 0 X-129 X-127 HO 0 H o OH OH H,, OH HO OH 0 0 DIsulfonly1 Halides -C F. Nit NOFH. 0 0 CIOO O 0 CH3H "IKO F- O0 CH3 X-13 0 138 X-13X- 01J2 X-3136 0 F07 0 0 0 0 CH3 O X- I 4.07 X-13 0 0 X-4 0-3 X-14 X- 1J7 o X-X-141 WO 01/42193 PCT/USOO/33057 == HjC 7 CH-J 0\ C/' HjC 1 1 0i~/ O S C 0 CI/I 0 0-4 HjC0 X-143 X- 142 0 C/ 0 F" N NN /0 0 NS, X-145 X- 146 0~/' 0 0C// I F / \ f Hj C 7 CHJ CN/ l Cl-f X- 148 HO00 X- 147 X- 149X15 0 11 IC/ 04,/ N X- 152 0 0 0 X- 15 1 Dioldehydes 01 X-50NH X- 154, CHJ 0K N'0. X- 156 \,o 0 7H 0 0 X- 15500N 0- CHl 3 X- 158 X- 157 WO 01/42193 PCT/USOO/33057 --92- HXC X115 0 00 N NN 0 X- 164 X- 161 0 X- 162CH 0~ Q 0 O N N N OH 0I-1511 X- 166 0 X-167 -0 HjcO 0 0 X- 169 X- 170 N OH '0 NO 0 H Ce- - 7

/

1 J X- 172 HO X- s174' X- 173 Nl"' ~ CHJi X- 177 Diholides

C-~~N

5 0 ~ X- 176 X- 175 Br B?~r Br Br I~N~IOH OH X- 178 X- 179 X- 180 -93- CI 0 y C Br Br X-182 X-181 X- 183 Br Br CH2 X- 184 X-185 Y-186 OH N r C X-187 Br Br X-188 Br Br Xr X-191 X-189 X-190 0 Br Br 0 BrB /-0 H C

CH

3 X193 3 X-194 F Br C iC HO OH Br F CH3I Br Br Br X-196 X-197 X-198 X 195 H 3 0 X-200

H

3 CH Br X-199 CX 201 Br OH Br 0//l-\l- lBr Br, C/Bral . -Br O Br Br X-205 X06 X-202 X-203 X204-20 Br Br CB Br

H

3 C-0

H

2 N Br X-209 X-210 c' - X-208 X-207 B B Ci N N Br CH3 X-213 X N214 X-211 X-212 z WO 01/42193 PCT/USOO/33057 Disocyanates 0 0 Na NN X-215 0 X-216 0 0 N 0 / 0 N N N0 HJC-O 0-C H 3 X-218 0 X-217F FF F F 0=-= N F /H 3 C N NNN X-219 X-220 0 X-221 /N N 0 Br C-j0 N N CiJ. X-222 HJC CHj X-224 X-223 N N N

-

N N N N26X-227 0 X-226 CH3 CjH3 N NN N 0 0 0 X-225 X-229 X-228 c/ CHJ N N N7 X-230 X-231 X-232 X-23

/

WO 01/42193 PCT/USOO/33057 . -95-- 0 * *00 00 0 N

H

3 Co CHN--' CHH, 0 -3 HX-23 X-2J4 H CH3j CHJ H3C CH3 Cl-I CCH N NN X-236 H 3 C CH 0 0 0 0 X-237 I~ CHH - X-238 H 3 H3C CH 3 0 0 -239 CH3 0 C CI X-240 N 0 N N /C 0 X-241 0~ X-242 0 CH=~- H N~ CH3 N CH3 0 0 H3C CH3 N N X-243 X-244 0 CH X-244 X-245. C3 WO 01/42193 PCT/USOO/33057 00 0 N N ~ 3 N N"~ N' I-jC 11 X 24 0 // X-246 0 X-247 Diom in es I ~X-249 ~N~~ ('NNN~ X-51 H 2 N NH 2 INJ N X-25252 X-250HN N x

CH

3 CHj 9 CH N HN OH NHH H 2 N NH 2 X-25 CA. ClJI C/-I K- N

H

2 NX-54 NH 2 X-256.

H

2 N~~0- NH 2

H

2 N N \* X-255 X-257

NH

2 N

N

2 H2N

~NH

2

H

2 N NH 2 X-260 X-258 X-259 0

NH

2 X-262 X-261

NH

2 '

H

2 N N H 2 J> NH2 N X-265 X-63 H 2 N X-264 WO 01/42193 PCT/USOO/33057 -97-

NH

2 N N dN OH X-267 OH X-266 HO. OH OH HO XH2N N C y H2N X-270 2 HOH3 N2 NH 2

H

2 N NH2 C3 Cq~j X-271 X-272 H3C N N H2N NH

CNH

2 3C CHJ

H

2 N

NH

2 S X-274X27 X-2757 CH3 CH32-7. XX2762

NH

2

NH

2 HO N ~ N OH X-278X-280 0 H2N

NH

2 /\ 0/

NH

2 N N \ NHN 0 NH2

H

2 N NH

NH

2 X-283X WO 01/42193 PCT/USOO/33057 -98-

H

2 N0O 0 NH2 X-285 HJC N N CH 3

H

2 N

NH

2 X-286 X-287 0 0 H2C H2N N CH 3 H2NI NH2 S N HN~~NHJC

CH

3 X-289 X-290 X-288 NH 2 X-292

H

2 N 0 0

NH

2 0 0 X-291 U0 2S NH

H

2 NX-97NH HC NNH3 HN NH2 X-298

H

2 N X-294 2

H

2 N

NH

2 0 . N0 X-295NH 2

NH

2 -X-295 0'N 0 X26 H 2 N X-29 7 NH 2 Hj~ N N---' ,.,N , CH 3

H-

2 N

NH

2 X-298 X-299

H

2 N

NH

2

H

2 N

NH

2

H

2 N

NH

2 X-301 X-302 X-300 WO 01/42193 PCT/USOO/33057 -99--. 0 0 a0 H2N NH2

H

2 N

NH

2 X-304 X-30J CHj H2N 0 o 0

NH

2 N H2N NH X-305

''N'CH

3 X-307 Chiral X-306

NH

2

NH

2 CHi CHi

H

2 N

NH

2 Cl-I CHJ X- 308 X-J09

NH

2 N2 CHJ H 2 NCIj H3 2 CHH2 (yN HjC

'CH

3

CH

3 N Chiral X-313

H

2 X-312 X-314 WO 01/42193 PCT/USOO/33057 100-C 3 H2 N N2 X-315 HjC NN X-316 HjC CH 3 CI CI C H3C N X-317 N N Chiral

CH

3 X-318 NH2

H

2 N N

NH

2 HN >K'N NH 2 X-319

H

3 C CH 3 X-320

H

3 C N N CH3 H 2 N ',- ' NH 2 X-321 X-322 H3 N NCH H3CN N CH 3

H

2 N NH2 X-323 X-324 X-325 Diols CHHO Br H3C Br HO OOH X-2 0O - 0 X-327 0 Br Br OH X-326 N OH 0 HO N N O 0 X-328 X-329

N

WO 01/42193 PCT/USOO/33057 o OH HO 1 OH N X-331 OH H.

CH

3 OH X-332 X- 330 00 0 OH r' O

H

3 C C0CH

HO~.

0 H 3 X-333 OH OH

H

3 C OH 0O HN O X-335 X- 336 X-37 HO O ~-~O O%*- O~ OH HO S OH X-338 X-339 F FF3 OH HHOF F H3C 0CH3 O HO X-343 X-340 OH-34 O HOH OH C

H

3 C CO-H X-JJ5 X-2 XJ ~HO 0 H H OH X-JJ8 X-344 WO 01/42193 PCT/USOO/33057 --102 HO S OH X-J45 HO OH X-346 0 0 OH HO OH Hj CH3 HO OH 6H X-347 X-J48 X-349 0 HOO ~OH OH HO 0 HO~ X-350 X-351 OH HO HO OH X- 352 X- 353 OH F F HO OH F F O F FH X-J55 5X-354 OH 0 0OHHO HO HX-357 \ \HO OH X-356 X-358 OH OH H C OH OH Hy X-359 X-360 WO 01/42193 PCT/USOO/33057 --103- OH OH HO HO O X-J61 X-362 H00 CH0H HO OH HO -- ' CH 2 HO.Sa.HO I OH HX- 365 OHi~ HO ) I~ X-363 .X-364 OH OH HO OH HjC CHj X-366 X-J67 HO 0 OH HO HO 0 X-3J68X-6 X-369 HO OOH OH X-J70 H3C CH 3 - X-371 OH X- 372 OH OH N O OH X-373 OH HOHO CHy

CH

3 X-375 X-374

OH

WO 01/42193 PCT/USOO/33057 --104- CHj OH HOOH H3C CH3 HO OH HHC OH HO X-377 X-378 X-376 CH3 CCH5 HO OH HO"O X-379 X-380 HO S OH HO OH HO OH X-381 X-382 X-383 F FF HO HO OH X-384FFF X-385 Dithiols HSSH HS SH C SH X-386 X-387 X-388 SH HS CHi 3 HSS H/S X-389 SH X-391 X-390 WO 01/42193 PCT/USOO/33057 -105-- HS 0 H HS SH HS X-392 X-394 SH CH 3 SH SH HS OH SH X-395 HjC

CH

3 X-396 SH X-397 HS-I HS HS OH HS SH X-398 X-399 X-400

CH.

3 0

CH

3 HS-\ 1HS SH HS SH X-403 SH X-402 X-401 0 0 HS S/I HS SH HS'- >N L \ / X-404 X-405 X-406 X 407 X-407 X-408 X-409 HSH H HS. OH OH SF OH X-410 X HS HS HS SH OH/ OH SH H Sr2 OH X410 X X-481 X-40 SH S OHOHS HS SHS HS SH OH SH X-41 X1 X-418 X-417 WO 01/42193 PCT/USOO/33057 --106- Representative ligands for use in this invention include, by way of example, L-1 and L-2 as identified above wherein L-l is selected from a compound of formula (a) and L-2 is selected from a compound of formula (b). Combinations of ligands (L) and linkers (X) per this invention include, by 5 way example only, homo- and hetero-dimers wherein a first ligand is selected from L-1 and the second ligand and linker is selected from the following: L-2/X-1- L-2/X-2- L-2/X-3- L-2/X-4- L-2/X-5- L-2/X-6 L-2/X-7- L-2/X-8- L-2/X-9- L-2/X-10- L-2/X-1 1- L-2/X-12 10 L-2/X-13- L-2/X-14- L-2/X-15- L-2/X-16- L-2/X-17- L-2/X-18 L-2/X-19- L-2/X-20- L-2/X-21- L-2/X-22- L-2/X-23- L-2/X-24 L-2/X-25- L-2/X-26- L-2/X-27- L-2/X-28- L-2/X-29- L-2/X-30 L-2/X-31- L-2/X-32- L-2/X-33- L-2/X-34- L-2/X-35- L-2/X-36 L-2/X-37- L-2/X-38- L-2/X-39- L-2/X-40- L-2/X-41- L-2/X-42 15 L-2/X-43- L-2/X-44- L-2/X-45- L-2/X-46- L-2/X-47- L-2/X-48 L-2/X-49- L-2/X-50- L-2/X-51- L-2/X-52- L-2/X-53- L-2/X-54 L-2/X-55- L-2/X-56- L-2/X-57- L-2/X-58- L-2/X-59- L-2/X-60 L-2/X-61- L-2/X-62- L-2/X-63- L-2/X-64- L-2/X-65- L-2/X-66 L-2/X-67- L-2/X-68- L-2/X-69- L-2/X-70- L-2/X-71- L-2/X-72 20 L-2/X-73- L-2/X-74- L-2/X-75- L-2/X-76- L-2/X-77- L-2/X-78 L-2/X-79- L-2/X-80- L-2/X-81- L-2/X-82- L-2/X-83- L-2/X-84 L-2/X-85- L-2/X-86- L-2/X-87- L-2/X-88- L-2/X-89- L-2/X-90 L-2/X-91- L-2/X-92- L-2/X-93- L-2/X-94- L-2/X-95- L-2/X-96 L-2/X-97- L-2/X-98- L-2/X-99- L-2/X-100- L-2/X-101- L-2/X-102 25 L-2/X-103- L-2/X-104- L-2/X-105- L-2/X-106- L-2/X-107- L-2/X-108 L-2/X-109- L-2/X-1 10- L-2/X-1 11- L-2/X- 112- L-2/X-113- L-2/X- 114 L-2/X-115- L-2/X-1 16- L-2/X-117- L-2/X-118- L-2/X-1 19- L-2/X-120 L-2/X- 121- L-2/X- 122- L-2/X-123- L-2/X- 124- L-2/X-125- L-2/X- 126 L-2/X-127- L-2/X-128- L-2/X- 129- L-2/X-130- L-2/X-131- L-2/X-132 30 L-2/X-133- L-2/X-134- L-2/X-135- L-2/X-136- L-2/X-137- L-2/X-138 L-2/X-139- L-2/X- 140- L-2/X- 141- L-2/X- 142- L-2/X-143- L-2/X- 144 L-2/X-145- L-2/X- 146- L-2/X- 147- L-2/X-148- L-2/X- 149- L-2/X-150 L-2/X- 151- L-2/X- 152- L-2/X-153- L-2/X-154- L-2/X-155- L-2/X- 156 L-2/X- 157- L-2/X-158- L-2/X-159- L-2/X- 160- L-2/X- 161- L-2/X- 162 35 L-2/X-163- L-2/X-164- L-2/X-165- L-2/X-166- L-2/X-167- L-2/X-168 L-2/X- 169- L-2/X- 170- L-2/X- 171- L-2/X-172 L-2/X-173- L-2/X- 174- L-2/X-175- L-2/X- 176- L-2/X- 177- L-2/X-178- WO 01/42193 PCT/USOO/33057 --107- L-2/X- 179- L-2/X- 180- L-2/X- 18 1- L-2/X- 182- L-2/X- 183- L-2/X- 184 L-2/X- 185- L-2/X-186- L-2/X- 187- L-2/X-188- L-2/X-189- L-2/X-190 L-2/X- 19 1- L-2/X- 192- L-2/X- 193- L-2/X- 194- L-2/X- 195- L-2/X- 196 L-2/X- 197- L-21X- 198- L-2/X- 199- L-2/X-200- L-2/X-20 1- L-2/X-202 5 L-2/X-203- L-2/X-204- L-2/X-205- L-2/X-206- L-2/X-207- L-2/X-208 L-2/X-209- L-2/X-2 10- L-2/X-21 1- L-2/X-212- L-2/X-213- L-2/X-214 L-2/X-2 15- L-2/X-2 16- L-2/X-2 17- L-2/X-2 18- L-2/X-2 19- L-2/X-220 L-2/X-221I- L-2/X-222- L-2/X-223- L-2/X-224- L-2/X-225- L-2/X-226 L-2/X-227- L-2/X-228- L-2/X-229- L-2/X-230- L-2/X-23 1 - L-2/X-232 10 L-2/X-233- L-2/X-234- L-2/X-235- L-2/X-236- L-2/X-237- L-2/X-238 L-2/X-239- L-2/X-240- L-2/X-241 - L-2/X-242- L-2/X-243- L-2/X-244 L-2/X-245- L-2/X-246- L-2/X-247- L-2/X-248- L-2/X-249- L-2/X-250 L-2/X-25 1 - L-2/X-252- L-2/X-253 - L-2/X-254- L-2/X-255- L-2/X-256 L-2/X-257- L-2/X-258- L-2/X-259- L-2/X-260- L-2/X-261 - L-2/X-262 15 L-2/X-263- L-2/X-264- L-2/X-265- L-2/X-266- L-2/X-267- L-2/X-26 8 L-2/X-269- L-2/X-270- L-2/X-271- L-2/X-272- L-2/X-273- L-2/X-274 L-2/X-275- L-2/X-276- L-2/X-277- L-2/X-278- L-2/X-279- L-2/X-280 L-2/X-281- L-2/X-282- L-2/X-283- L-2/X-284- L-2/X-285- L-2/X-286 L-2/X-287- L-2/X-288- L-2/X-289- L-2/X-290- L-2/X-291I- L-2/X-292 20 L-2/X-293- L-2/X-294- L-2/X-295- L-2/X-296- L-2/X-297- L-2/X-298 L-2/X-299- L-21X-300- L-2/X-301- L-2/X-302- L-2/X-303- L-2/X-304 L-2/X-305- L-2/X-306- L-2/X-307- L-2/X-308- L-2/X-309- L-21X-3 10 L-2/X-31I1I- L-2/X-312- L-2/X-313- L-2/X-314- L-2/X-315- L-2/X-316 L-2/X-3 17- L-2/X-3 18- L-2/X-3 19- L-2/X-320- L-2/X-32 1 - L-2/X-322 25 L-2/X-323- L-2/X-324- L-2/X-325- L-2/X-326- L-2/X-327- L-2/X-328 L-2/X-329- L-2/X-330- L-2/X-33 1- L-2/X-332- L-2/X-3 33- L-2/X-3 34 L-2/X-33 5- L-2/X-336- L-2/X-33 7- L-2/X-33 8- L-2/X-3 39- L-2/X-340 L-2/X-341- L-2/X-342- L-2/X-343- L-2/X-344- L-2/X-345- L-2/X-346 L-2/X-347- L-2/X-348- L-2/X-349- L-2/X-350- L-2/X-351- L-2/X-352 30 L-2/X-353- L-2/X-354- L-2/X-355- L-2/X-356- L-2/X-357- L-2/X-358 L-2/X-359- L-2/X-360- L-2/X-361- L-2/X-362- L-2/X-363- L-2/X-364 L-2/X-365- L-2/X-366- L-2/X-367- L-2/X-368- L-2fX-369- L-2/X-370 L-2/X-371- L-2/X-372- L-2/X-373- L-2/X-374- L-2/X-375- L-2/X-376 L-2/X-3 77- L-2/X-3 78- L-2/X-3 79- L-2/X-3 80- L-2/X-3 81- L-2/X-3 82 35 L-2/X-383- L-2/X-384- L-2/X-385- L-21X-386- L-2/X-387- L-2/X-388 L-2/X-3 89- L-2/X-390- L-2/X-391- L-2/X-392- L-2/X-393- L-2/X-394 L-2/X-3 95- L-2/X-396- L-2/X-397- L-2/X-398- L-2/X-399- L-2/X-400 L-2/X-401I- L-2/X-402- L-2/X-403- L-2/X-404- L-2/X-405- L-2/X-406- WO 01/42193 PCT/USOO/33057 --108- L-2/X-407- L-2/X-408- L-2/X-409- L-2/X-410- L-2/X-411- L-2/X-412 L-2/X-4 13- L-2/X-4 14- L-2/X-4 15- L-2/X-4 16- L-2/X-4 17- L-2/X-4 18 and so on. 5 Utility, Testing, and Administration Utility The multibinding compounds of this invention are P2 adrenergic receptor agonists or partial agonists. Accordingly, the multibinding compounds and pharmaceutical compositions of this invention are useful in the treatment and 10 prevention of diseases mediated by P2 adrenergic receptor such as asthma, chronic obstructive pulmonary disease, bronchitis, and the like. They are also useful in the treatment of nervous system injury and premature labor. It is also contemplated that the compounds of this invention are useful for treating metabolic disorders such as obesity, diabetes, and the like. 15 Testing The P2 adrenergic receptor agonistic activity of the compounds of Formula (I) to may be demonstrated by a variety of in vitro assays known to those of ordinary skill in the art, such as the assay described in the biological Examples 1 20 and 2. It may also be assayed by the ex vivo assays described in Ball, D. I. et al., "Salmeterol a Novel, Long-acting beta 2-Adrenergic Agonist: Characterization of Pharmacological Activity in Vitro and in Vivo" Br. J. Pharmacol., 104, 665-671 (1991); Linden, A. et al., "Salmeterol, Formoterol, and Salbutamol in the Isolated Guinea-Pig Trachea: Differences in Maximum Relaxant Effect and Potency but not 25 in Functional Atagonism. Thorax, 48, 547-553, (1993); and Bials, A. T. et al., Investigations into Factors Determining the Duration of Action of the Beta 2 Adrenoceptor Agonist, Salmeterol. Br. J. Pharmacol., 108, 505-515 (1993); or in vivo assays such as those described in Ball, D. I. et al., "Salmeterol a Novel, Long acting beta 2-Adrenergic Agonist: Characterization of Pharmacological Activity in 30 Vitro and in Vivo" Br. J. Pharmacol., 104, 665-671 (1991); Kikkawa, H. et al., "TA-2005, a Novel, Long-acting, and Selective Beta 2-Adrenoceptor Agonist: Characterization of its in vivo Bronchodilating Action in Guinea Pigs and Cats in WO 01/42193 PCT/USOO/33057 --109- Comparison with other Beta 2-Agonists". Biol. Pharm. Bull., 17, 1047-1052, (1994); and Anderson, G. P., "Formoterol: Pharmacology, Molecular basis of Agonism and Mechanism of Long Duration of a Highly Potent and Selective Beta 2-Adrenoceptor Agonist Bronchodilator, Life Sciences, 52, 2145-2160, (1993). 5 Pharmaceutical Formulations When employed as pharmaceuticals, the compounds of this invention are usually administered in the form of pharmaceutical compositions. These compounds can be administered by a variety of routes including oral, rectal, 10 transdermal, subcutaneous, intravenous, intramuscular, and inhalation (e.g., intranasal or oral inhalation). These compounds are effective as injectable, inhaled and oral compositions. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one active compound. A preferred manner for administering compounds of this invention is inhalation. This is an 15 effective means for delivering a therapeutic agent directly to the respiratory tract for the treatment of diseases such as asthma and other similar or related respiratory tract disorders (see U. S. Patent No. 5,607,915). This invention also includes pharmaceutical compositions which contain, as the active ingredient, one or more of the compounds described herein associated 20 with pharmaceutically acceptable carriers. In making the compositions of this invention, the active ingredient is usually mixed with an excipient, diluted by an excipient or enclosed within such a carrier which can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for 25 the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders. 30 In preparing a formulation, it may be necessary to mill the active compound to provide the appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it ordinarily is milled WO 01/42193 PCT/USOO/33057 --1 10to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size is normally adjusted by milling to provide a substantially uniform distribution in the formulation, e.g. about 40 mesh. Some examples of suitable excipients include lactose, dextrose, sucrose, 5 sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. The formulations can additionally include: lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as 10 methyl- and propylhydroxy-benzoates; sweetening agents; and flavoring agents. The compositions of the invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art. The compositions are preferably formulated in a unit dosage form. The term 15 "unit dosage forms" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Preferably, the compound of Formula (I) above is employed at no more than about 20 weight percent of the pharmaceutical 20 composition, more preferably no more than about 15 weight percent, with the balance being pharmaceutically inert carrier(s). The active compound is effective over a wide dosage range and is generally administered in a pharmaceutically effective amount. For example, when the drug is administered via inhalation, each dosage contains from about 1 Ig to about 1000 25 pg, preferably about 2 pg to about 500 pg, more preferably about 5 jig to about 100 pg, even more preferably about 5 pg to about 60 pg, of the active ingredient. It, will be understood, however, that the amount of the compound actually administered will be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of 30 administration, the actual compound administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like. Furthermore, the compound of this invention may be WO 01/42193 PCT/USOO/33057 --111 administered prophylactically, for example, a pharmaceutical composition containing a compound of this invention may be administered before the bronchospasm begins in an asthma attack, to prevent its occurrence or to reduce the extent to which it occurs. 5 For preparing solid compositions such as tablets, the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention. When referring to these pre-formulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition 10 so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. This solid pre-formulation is then subdivided into unit dosage forms of the type described above containing the active ingredient of the present invention. The tablets or pills of the present invention may be coated or otherwise 15 compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permit the inner component to pass intact into the 20 duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate. The liquid forms in which the novel compositions of the present invention 25 may be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils such as corn oil, cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles. Compositions for inhalation or insufflation include solutions and 30 suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders (see U. S. Patent No. 5,983,956). The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as WO 01/42193 PCT/USOO/33057 --112described supra. Preferably the compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the 5 nebulizing device may be attached to a face mask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices which deliver the formulation in an appropriate manner (see U. S. Patents Nos. 5,919,827 and 5,972,919). 10 Furthermore, the pharmaceutical compositions containing one or more compound(s) of this invention can be administered in combination any other suitable drug, for example, with a suitable steroidal anti-inflammatory drug, e.g., budesonide, flucatisone, beclamethasone, for the treatment of respiratory disorders. When the combination therapy is employed, the pharmaceutical composition 15 containing the compound(s) of this invention and the steroidal anti-inflammatory drug may be administered simultaneously, sequentially or separately. Each component used in the combination therapy is employed in an amount sufficient for its intended purpose. For example, the steriodal anti-inflammatory drugs are employed in sufficient amounts to effect reduction in inflammation in vivo. The p 20 2 adrenergic receptor agonist/partial agonist compounds of this invention are employed in an amount sufficient to cause relaxation of smooth muscle tissue, for example, in the bronchial system. Preferably, the dose range for compounds of this invention is from about 1 ptg to about 1000 ptg per dose, more preferably about 2 pg to about 500 pg, even 25 more preferably about 5 pg to about 100 pg, and still more preferably about 5 pig to about 60 ig. The preferred dosage range for a steroidal anti-inflammatory drug is from about 50 to 4800 [tg and more preferably about 100 pg to about 1600 Ig. Again, the particular dose used will depend on the patient (age, weight, etc.), and the severity of the disease (mild, moderate, severe). Lastly, a pharmaceutical 30 composition containing the two active ingredients can also be prepared for administering the drugs simultaneously.

WO 01/42193 PCT/USOO/33057 --113- EXAMPLES The following preparations and examples are given to enable those skilled in the art to more clearly understand and to practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being 5 illustrative and representative thereof. In the examples below, the following abbreviations have the following meanings. Unless otherwise stated, all temperatures are in degrees Celsius. If an abbreviation is not defined, it has its generally accepted meaning. 10 A = Angstroms cm = centimeter DCC = dicyclohexyl carbodiimide DMF = N,N-dimethylformamide DMSO = dimethylsulfoxide 15 g = gram HPLC = high performance liquid chromatography MEM = minimal essential medium mg = milligram MIC = minimum inhibitory concentration 20 min minute mL = milliliter mm = millimeter mmol = millimol N = normal 25 THF = tetrahydrofuran pL= microliters pIm = microns rt = room temperature Rf = retention faction 30 NMR = nuclear magnetic resonance ESMS = electrospray mass spectrum ppm = parts per million 35 Synthetic Examples Example 1 WO 01/42193 PCT/USOO/33057 --1 14- Synthesis of trans-1,4-bis {N-[2-(4-hydroxy-3-hydroxymethylphenyl)-2 hydroxyethyl]amino} cyclohexane (following figure 5) OH H OH H H N. N H HO OH HO OH Step 1 5 To a solution of 5-acetylsalicylic acid methyl ester 11 (5.0g, 25.7 mmole) in dimethylsulfoxide (44 mL) was added 48% hydrobromic acid. The resulting mixture was stirred at 55 'C for 24 h, and poured into a slurry of ice-water (200 mL), precipitating a pale yellow solid. The solid was filtered, washed with water (200 mL), and dried to give c,a-dihydroxy-4-hydroxy-3-methoxycarbonyl 10 acetophenone 12. The product was re-suspended in ethyl ether (200 mL), filtered and dried to give (3.41 g, 59%) of pure product. R 1 = 0.8 (10% MeOH/CH 2 Cl 2

)

H

1 -NMR (4/1 CDCl 3

/CD

3 0D, 299.96 MHz): 8 (ppm) 8.73-8.72 (d, 1H), 8.28 8.24 (dd, 1H), 7.08-7.05 (d, 1H), 5.82 (s, 1H), 4.01 (s, 3H). Step 2 15 To a suspension of x,c-dihydroxy-4-hydroxy-3-methoxycarbonyl acetophenone 12 (0.3 g, 1.33 mmole) in THF (10 mL) was added a solution of trans-1,4-diaminocyclohexane (76 mg, 0.66 mmole) in THF (5 mL). The resulting suspension was stirred for 3 h at ambient temperature under nitrogen atmosphere, at which formation of an imine was completed judged by TLC analysis. After cooling 20 of the resulting solution at ice bath, an excess amount of 2M BH 3 -Me 2 S in hexane (4 mL, 8 mmole) was added to the previous solution. The resulting mixture was slowly warmed to rt and refluxed for 4 h under N 2 stream. After cooling the reaction mixture, MeOH (5 mL) was added to quench excess amount of 2M BH 3 Me 2 S. After stirring for 30 min., the final solution (or cloudy solution) was 25 evaporated in vacuo, yielding a pale brown solid. The solid was washed with EtOAc/hexane (1/2; 20 mL), and dried. The crude product was dissolved in 50% MeCN/H 2 0 containing 0.5% TFA, and purified by prep-scale high performance WO 01/42193 PCT/USOO/33057 --1 15liquid chromatography (HPLC) using a linear gradient (5% to 50% MeCN/H 2 0 over 50 min, 20 mL/min; detection at 254 nM). Fractions with UV absorption were analyzed by LC-MS to isolate trans-1,4-bis{N-[2-(4-hydroxy-3-hydroxymethyl phenyl)-2-hydroxyethyl]amino} cyclohexane 13. 5 Hi1-NMR (CD 3 0D, 299.96 MHz): 6 (ppm) 7.35 (d, 2H), 7.18 (dd, 2H), 6.80 6.78 (d, 2H), 4.88-4.86 (m, 2H), 4.65 (s, 4H), 3.15 (br s, 4H), 2.89 (m, 2H), 1.68 1.55 (br m, 4H); ESMS (C 2 4

H

3 4

N

2 0 6 ): calcd. 446.5, obsd. 447.5 [M+H]*. Compound 14: Proceeding as described above but substituting trans-1,4-diamino 10 cyclohexane with 4,4'-methylenebis(cyclohexylamine) gave bis{4,4'-[N-[2-(4 hydroxy-3 -hydroxymethylphenyl)-2-hydroxyethyl]amino]cyclohexane} methane. ESMS (C 3 1

H

4 6

N

2 0 6 ): calcd. 542.7, obsd. 543.6 [M+H]*. Compound 15: Proceeding as described above but substituting trans-1,4-diamino 15 cyclohexane with 1,3-cyclohexanebis(methylamine) gave 1,3-bis{N-[2-(4-hydroxy 3-hydroxymethylphenyl)-2-hydroxyethyl]aminomethyl}cyclohexane. ESMS (C 27

H

3 8

N

2 0 6 ): calcd. 474.6, obsd. 475.3 [M+H]*. Compound 16: Proceeding as described above but substituting trans-1,4-diamino 20 cyclohexane with 1,8-diamino-p-menthane gave 1,8-bis{N-[2-(4-hydroxy-3 hydroxymethylphenyl)-2-hydroxyethyl]amino } -p-menthane. ESMS (C 28

H

4 2

N

2 0 6 ): calcd. 502.6, obsd. 503.3 [M+H]*. Compound 17: Proceeding as described above but substituting trans-1,4-diamino 25 cyclohexane with 1,4-bis(3-aminopropyl)piperazine gave 1,4-bis{3-[[N-2-(4 hydroxy-3-hydroxymethylphenyl)-2-hydroxyethyl]amino]propyl}piperazine. ESMS (C 28

H

4 4

N

4 0 6 ): calcd. 532.6, obsd. 533.3 [M+H]*, 555.0 [M+Na]*. Compound 18: Proceeding as described above but substituting trans-1,4-diamino 30 cyclohexane with p-xylylenediamine gave 1,4-bis {N- [2-(4-hydroxy-3 -hydroxy methylphenyl)-2-hydroxyethyl]aminomethyl} benzene. ESMS (C 6

H

3 2

N

2 0 6 ): calcd. 468.5, obsd. 469.3 [M+H]*, 492.0 [M+Na]*.

WO 01/42193 PCT/USOO/33057 --1 16- Compound 19: Proceeding as described above but substituting trans-1,4-diamino cyclohexane with m-xylylenediamine gave 1,3-bis{N-[2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl] aminomethyl } benzene. ESMS (qH 3 2

N

2 0 6 ): calcd. 5 468.5, obsd. 469.3 [M+H]*, 492.0 [M+Na]*. Compound 20: Proceeding as described above but substituting trans-1,4-diamino cyclohexane with 2-aminobenzylamine gave 1- {N-[2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl]aminomethyl} -2- {N- [2-(4-hydroxy-3 10 hydroxymethylphenyl)-2-hydroxyethyl]amino }benzene. ESMS (C 5

H

30

N

2 0 6 ): calcd. 454.5, obsd. 455.3 [M+H]*. Compound 21: Proceeding as described above but substituting trans-1,4-diamino cyclohexane with 2-(4-aminophenyl)ethylamine gave 1- {2- [N-2- [(4-hydroxy-3 15 hydroxymethylphenyl)-2-hydroxyethyl]amino]ethyl}-2-{N-[2-(4-hydroxy-3 hydroxymethylphenyl)-2-hydroxyethyl] amino]benzene. ESMS (q 6

H

32

N

2 0 6 ): calcd. 468.5, obsd. 469.3 [M+H]*. Compound 22: Proceeding as described above but substituting trans-1,4-diamino 20 cyclohexane with 4,4'-oxydianiline gave 4,4'-bis{N-[2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl]amino } phenylether. ESMS (qH 3 2

N

2 0 7 ): calcd. 532.6, obsd. 533.3 [M+H]*, 556.1 [M+Na]*. Compound 23: Proceeding as described above but substituting trans-1,4-diamino 25 cyclohexane with 2-aminobenzylamine gave I -{N-[2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl]aminomethyl}-4-{N-[2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl]amino} benzene. ESMS (q 5

H

3 0

N

2 0 6 ): calcd. 454.5, obsd. 455.5 [M+H]*, 477.3 [M+Na]*. 30 Example 2 Synthesis of 1-{2-[N-2-[(4-hydroxy-3-hydroxymethylphenyl)-2 hydroxyethyl]amino]ethyl} -4- {N-[2-phenyl-2-hydroxyethyl]amino]benzene WO 01/42193 PCT/USOO/33057 --117- (following figure 6) OHH OH H 0 N N HO O H O To a suspension of c,ac-dihydroxy-4-hydroxy-3-methoxycarbonyl 5 acetophenone 12, prepared in Example 1, Step 1 above, (0.3 g, 1.33 mmole) in THF (10 mL) was added a solution of 2-(4-aminophenyl)ethylamine 25 (0.181 g, 1.33 mmol) in THF (5 mL). The resulting suspension was stirred for 3 h at ambient temperature under nitrogen atmosphere, followed by addition a,c-dihydroxy acetophenone 24 (0.2g, 1.32 mmole). The reaction mixture was stirred for 3 h at 10 RT, at which formation of the imine was completed as judged by TLC analysis. The reaction mixture was cooled in an ice bath and an excess amount of 2M BH 3 Me 2 S in hexane (9 mL; 18 mmole) was added. The resulting mixture was slowly warmed to rt, and refluxed for 4 h under N 2 stream. After cooling, MeOH (10 mL) was added to quench excess amount of BH 3 -Me 2 S. After stirring 30 min., at rt, the 15 final solution (or cloudy suspension) was evaporated in vacuo, to give a pale brown solid. The solid was washed with EtOAc/hexane (1/2; 20 mL), and dried. The crude product was dissolved in 50% MeCN/H 2 0 containing 0.5% TFA, and purified by prep-scale high performance liquid chromatography (HPLC) using a linear gradient (5% to 50% MeCN/H 2 0 over 50 min, 20 mL/min; detection at 254 20 nM). Fractions with UV absorption were analyzed by LC-MS to locate 1-{2-[N-2 [(4-hydroxy-3 -hydroxymethylphenyl)-2-hydroxyethyl] amino] -ethyl} -4- {N-[2 phenyl-2-hydroxyethyl]amino]benzene 26. ESMS (C 5

H

3 0

N

2 0 4 ): calcd. 422.5, obsd. 423.3 [M+H]*. Compound 27: 25 Proceeding as described above, but substituting c,ax-dihydroxy-4-hydroxy 3 -methoxycarbonylacetophenone with a,c-dihydroxyacetophenone gave 1- {2- [N- WO 01/42193 PCT/USOO/33057 --118- [2-phenyl-2-hydroxyethyl]aminoethyl} -4-[N-(2-phenyl-2-hydroxyethyl)amino] benzene. ESMS (C 2 4

H

2 8

N

2 0 8 ): calcd. 376.5, obsd. 377.0 [M+H]*. Example 3 5 Synthesis of 1-{2-[N-2-(4-hydroxy-3-hydroxymethylphenyl)-2-hydroxyethyl] amino]ethyl} -4- [N-(2-phenyl-2-hydroxyethyl)amino]benzene (following figure 7) OH H HO N HO H OH Step 1 To a solution of 4-(2-aminoethyl)aniline 25 (20 g, 147 mmole) in methanol 10 (250 mL) was added (Boc) 2 0 (32.4 g, 148 mmole) in methanol (50 mL) at rt. After stirring for 24 h, the reaction mixture was concentrated to dryness to afford a pale yellow oily residue. The oily material solidified slowly; thus it was dissolved in 5% MeOH/CH 2 Cl2, and subsequently applied to flash silica column chromatography (3 to 10% MeOH/CH 2 Cl 2 ). After purification, 4-(N-Boc-2-aminoethyl)aniline 28 was 15 obtained as a pale yellow solid (32.95g, 95%): Rf= 0.6 in 10% MeOH/CH 2 Cl 2

.

1

H

NMR (CD 3 0D, 299.96 MHz): 6 (ppm) 6.96-6.93 (d, 2H), 6.69-6.65 (d, 2H), 3.20 3.13 (q, 2H), 2.63-2.58 (t, 2H), 1.41 (s, 9H). Step 2 4-(N-Boc-2-aminoethyl)aniline 28 (1.25 g, 5.29 mmole) was dissolved in 20 methanol (30 mL), followed by addition of phenyl glyoxal 24 (0.708 g, 5.28 mmole). The reaction mixture was stirred for 1 h at rt, prior to addition of NaCNBH 3 (0.665 g, 10.6 mmole). The final mixture was stirred for 12 h at rt, concentrated, and purified by flash silica column chromatography (2 to 5% MeOH/CH 2 Cl2) to give N-(2-phenyl-2-hydroxyethyl)-4-(N-Boc-2-aminoethyl) 25 aniline as a pale yellow oil (1.71 g, 91%): Rf= 0.18 in 5% MeOH/CH 2 Cl 2 . 'H NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.4-7.25 (m, 5H), 7.0-6.95 (d, 2H), 6.63- WO 01/42193 PCT/USOO/33057 --1 19- 6.60 (d, 2H), 4.85-4.79 (dd, 1H), 3.3-3.21 (t, 2H), 3.2-3.15 (m, 2H), 2.64-2.5 (t, 2H), 1.42 (s, 9H). Step 3 A solution of N-(2-phenyl-2-hydroxyethyl)-4-(N-Boc-2-aminoethyl)aniline 5 (1.7 g, 4.77 mmole) in methylene chloride (10 mL) was cooled in ice bath, and TFA (10 mL) was slowly added under a stream of nitrogen gas. The reaction mixture was stirred for 1 h, and concentrated to yield a pale yellow oil. The crude material was purified by reversed phase HPLC (10% to 40% MeCN/H 2 0 over 50 min; 20 mL/min) to give N-(2-phenyl-2-hydroxyethyl)-4-(2-aminoethyl)aniline 29 as the 10 TFA salt (1.1 g). 1 H-NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.42-7.3 (in, 5H), 7.29-7.25 (d, 2H), 7.12-7.0 (d, 2H), 4.85-4.82 (in, 1H), 3.45-3.35 (in, 2H), 3.18-3.1 (t, 2H), 2.98-2.94 (t, 2H); ESMS (C 16

H

2 0

N

2 0 1 ): calcd. 256.4, obsd. 257.1 [M+H]*, 278.8 [M+Na]*, 513.4 [2M+H]*. Step 4 15 To a solution of N-(2-phenyl-2-hydroxyethyl)-4-(2-aminoethyl)aniline trifluoroacetate salt 29 (1.1 g, 2.3 mmole) in methanol (10 mL) was added 5 M NaOH solution (0.93 mL). After stirring for 10 min., the solution was concentrated to dryness. The residue was dissolved in THF (25 mL), and ac,ca-dihydroxy-4 hydroxy-3-methoxy-carbonylacetophenone 12 (0.514 g, 2.27 mmole) was added. 20 The reaction mixture was stirred for 12 h at rt, cooled to 0 'C, and BH 3 /Me 2 S (1.14 mL, 10 M ) was added under nitrogen atmosphere. The reaction mixture was gradually warmed to rt, stirred for 2 h at rt, and refluxed for 4 h. The reaction mixture was cooled and methanol (10 mL) was added slowly. After stirring for 30 min., at rt, the reaction mixture was concentrated to afford a solid residue, which 25 was dissolved in MeOH (20 mL) containing 10% TFA. Evaporation of the organics yielded a pale yellow oil which was purified by reversed phase HPLC: 10% to 30% MeCN/H 2 0 over 50 min; 20 mL/min to give 1-{2-[N-2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl]-amino] ethyl } -4- [N-(2-phenyl-2-hydroxyethyl) amino]benzene 30 as the TFA salt (0.65 g). 'H-NMR (CD 3 0D, 299.96 MHz): 6 30 (ppm) 7.42-7.3 (in, 6H), 7.28-7.24 (d, 2H), 7.18-7.14 (dd, 1H), 7.1-7.07 (d, 2H), 6.80-6.77 (d, 1H), 4.86-4.82 (in, 2H), 4.65 (s, 2H), 3.44-3.34 (in, 2H), 3.28-3.22 (in, WO 01/42193 PCT/USOO/33057 --120- 2H), 3.20-3.14 (in, 2H), 3.04-2.96 (in, 2H); ESMS (C 2 5

H

30

N

2 0 4 ): calcd. 422.5, obsd. 423.1 [M+H]*, 404.7 [M-1H 2 0]*, 387.1 [M-2H 2 0]*. Example 4 5 Synthesis of 1-{2-[N-2-(4-hydroxy-3-hydroxymethylphenyl)-2-hydroxyethyl] aminoethyl } -4- [N-(2-phenyl-2-(S)-hydroxyethyl)amino]benzene (following figure 8) OH H N HO N H HO OH Step 1 10 A solution of 4-(N-Boc-2-aminoethyl)aniline 28 (7.0 g, 29.6 mmole) in ethanol (100 mL) and (R)-styreneoxide (3.56 g, 29.6 mmole) was refluxed for 24 h. The organics were removed to give a pale yellow solid. N-(2-phenyl-2-(S) hydroxyethyl)-4-(N-Boc-2-aminoethyl)aniline was separated by flash silica column chromatography: 1/2 EtOAc/hexane to 3/1 EtOAc/hexane to 3% MeOH in 3/1 15 EtOAc/hexane: Rf= 0.39 in 3% MeOH/CH2Cl 2 Step 2 A solution of N-(2-phenyl-2-(S)-hydroxyethyl)-4-(N-Boc-2-aminoethyl) aniline (2.5 g, 7.0 mmole) in CH 2 Cl 2 (15 mL) was cooled in an ice bath under stream of nitrogen and TFA (15 mL) was slowly added. The reaction mixture was 20 stirred for 2 h at 0 0 C and then concentrated in vacuo. The crude product was dissolved in 20% MeCN/H 2 0 and purified by preparative reversed phase HPLC (5 to 2% MeCN/H 2 0 over 50 min; 254 nm; 20 mL/min.), to give N-(2-phenyl-2-(S) hydroxyethyl)-4-(2-aminoethyl)aniline trifluoroacetate salt 31 as a colorless oil. 'H NMR (CD 3 0D, 299.96 MHz): 8 (ppm); 7.45-7.25 (in, 9H), 4.9 (dd, 1H), 3.55-3.45 25 (in, 2H), 3.21-3.15 (t, 2H), 3.05-2.95 (t, 2H) ESMS (CIH 2

N

2 0): caled. 256.4, obsd. 257.1 [M+H]*, 280.2 [M+Na]*. Step 3 WO 01/42193 PCT/USOO/33057 --121- To a solution of N-(2-phenyl-2-(S)-hydroxyethyl)-4-(2-aminoethyl)aniline trifluoroacetate 31 (0.144 g, 0.3 mmole) in methanol (10 mL) was added aq. NaOH solution (1.0 M, 0.625 mL). The solution was concentrated to dryness and the residue was dissolved in anhydrous THF (5 mL). a,c-Dihydroxy-4-hydroxy-3 5 methoxycarbonylacetophenone 12 (0.067 g, 0.3 mmole) was added and the reaction mixture was stirred for 12 h at rt. BH 3 -Me 2 S (0.2 mL, 2M) was added at 0 0 C and the reaction mixture was heated at 75 *C for 6 h. After cooling the reaction mixture in ice bath, MeOH (5 mL) was slowly added to it to quench the reaction, and the reaction mixture was stirred for 30 min., at rt. The organics were removed and the 10 residue was dissolved in TFA/MeOH (1/9; 20 mL), and concentrated. The crude product was dissolved in 20% MeCN/H 2 0, and purified by preparative HPLC: 5 to 20% MeCN/H 2 0; 20 mL/min; 254 nm. ) to give 1-{2-[N-2-(4-hydroxy-3-hydroxy methylphenyl)-2-hydroxyethyl]amino]ethyl} -4-[N-(2-phenyl-2-(S)-hydroxyethyl) amino]benzene 33. 15 'H-NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.42-7.29 (m, 8H), 7.22-7.18 (d, 2H), 7.17-7.14 (dd, IH), 6.80-6.77 (d, 1H), 4.9-4.85 (m, 2H), 4.65 (s, 2H), 3.5-3.34 (m, 2H), 3.28-3.25 (m, 2H), 3.19-3.14 (m, 2H), 3.04-2.98 (m, 2H); ESMS

(C

25 H3 0

N

2 0 4 ): calcd. 422.5, obsd. 423.1 [M+H]*, 446.1 [M+Na]*. Proceeding as described in Example 4 above but substituting (R) 20 styreneoxide with (S)-styreneoxide gave 1-{2-[N-2-(4-hydroxy-3 hydroxymethylphenyl)-2-hydroxyethyl] amino]ethyl } -4-[N-(2-phenyl-2-(R) hydroxyethyl)amino]benzene 34. 1 H-NMR (CD 3 0D, 299.96 MHz): S (ppm) 7.42-7.28 (m, 8H), 7.20-7.1 (m, 3H), 6.80-6.77 (d, 1H), 4.9-4.85 (m, 2H), 4.65 (s, 2H), 3.45-3.34 (m, 2H), 3.28-3.25 25 (m, 2H), 3.19-3.15 (m, 2H), 3.04-2.98 (m, 2H); ESMS (C 25

H

30

N

2 0 4 ): calcd. 422.5, obsd. 423.1 [M+H]*, 446.1 [M+Na]*. Example 5 Synthesis of 1,6-bis{4-(N-[2-(4-hydroxy-3-hydroxymethylphenyl)-2 30 hydroxyethyl]aminohexyloxypropyl]phenoxy}hexane (following figure 9) WO 01/42193 PCT/USOO/33057 --122- OH

N-(CH

2

)

6 -0-(CH 2

)

3 O HO HO OH

N-(CH

2

)

6

-O-(CH

2

)

3 O HO HO Step 1 A solution of 3-(4-hydroxyphenyl)-1-propanol 35 (3.3 g, 21.7 mmole) and 1,6-di-iodohexane (3.5 g, 8.88 mmole) in dimethylsulfoxide (40 mL) was degassed and saturated with N 2 gas and potassium carbonate (4.5 g, 32.56 mmole) was added. 5 The reaction mixture was stirred at 80 'C for 18 h under nitrogen atmosphere and then quenched with brine (150 mL). The product was extracted with EtOAc (200 mL) and the organic extracts were washed with 0.1 M NaOH and brine, and dried with MgSO 4 . The organics were removed in vacuo to give a pale brown solid. The solid was purified by flash silica column chromatography: 4/1 hexane/EtOAc to 5% 10 MeOH in 1/1 hexane/EtOAc to give 1,6-bis[4-(3-hydroxypropyl)phenoxy]hexane 36 (Rf = 0.17 in 1/1 hexane/EtOAc) in 65% yield (2.23 g). 'H-NMR (CD 3 0D, 299.96 MHz): 5 (ppm) 7.08-7.05 (d, 4H), 6.80-6.77 (d, 4H), 3.93-3.89 (t, 4H), 3.56 3.52 (t, 4H), 2.64-2.56 (t, 4H), 1.81-1.69 (in, 8H), 1.44-1.21 (in, 4H). Step 2 15 A solution of 1,6-bis[4-(3-hydroxypropyl)phenoxy]hexane 36 (2.2 g, 5.69 mmole) in DMF (10 mL) was added to a solution of DMF (40 mL) containing NaH (0.57 g; 60% dispersion in mineral oil) at 0 "C under nitrogen atmosphere and the reaction mixture was heated at 50 "C. After 1 h, 6-bromohexanenitrile (2.26 mL, 17 mmole) was added and the reaction mixture was heated at 80 "C for 24 h. The 20 reaction mixture was quenched with brine solution (100 mL) and was extracted with EtOAc (250 mL). The organic phase was washed with brine, dried with MgSO 4 , and evaporated in vacuo, to give a pale yellow oil. Purification by flash silica column chromatography: 4/1 to 1/1 hexane/EtOAc afforded 1,6-bis[4-(5- WO 01/42193 PCT/USOO/33057 --123cyanopentyloxypropyl)]phenoxy]hexane 37 product (Rf = 0.6 in 1/1 EtOAc/hexane). 'H-NMR (CDCl 3 , 299.96 MHz): 8 (ppm) 7.09-7.06 (d, 4H), 6.82 6.79 (d, 4H), 3.94-3.90 (t, 4H), 3.42-3.37 (m, 8H), 2.64-2.58 (t, 4H), 2.40-2.32 (m, 8H), 1.90-1.26 (m, 24H). 5 Step 3 The 1,6-bis[4-(5-cyanopentyloxypropyl)]phenoxy]hexane 37 (0.278 g, 0.48 mmole) obtained in Step 2 above was added to a mixture of conc. HCl (10 mL) and AcOH (2 mL) and the reaction mixture was heated at 90 'C. After 15 h, the reaction mixture was diluted with brine (50 mL), extracted with EtOAc (100 mL), 10 and dried with MgSO 4 . Evaporation of the organic phase afforded the 1,6-bis[4-(5 carboxypentyl-oxypropyl)]phenoxy]hexane 38 as a pale yellow oily residue, which was used in next step without further purification. 'H-NMR (CDCl 3 , 299.96 MHz): 8 (ppm) 7.09-7.07 (d, 4H), 6.82-6.79 (d, 4H), 3.96-3.92 (t, 4H), 3.42-3.56 (m, 8H), 2.64-2.59 (t, 4H), 2.39-2.32 (m, 4H), 1.91-1.40 (m, 24H). 15 Step 4 To a solution of 2-hydroxy-2-(4-benzyloxy-3-hydroxymethylphenyl) ethylamine 39 (0.263 g, 0.96 mmole) in DMF (8 mL) was added 1,6-bis[4-(5 carboxypentyloxypropyl)phenoxy]hexane (~0.48 mmole), obtained in Step 3 above, HOBt (0.13 g, 0.96 mmole), DIPEA (0.21 mL, 1.20 mmole), and PyBOP (0.502 g, 20 0.96 mmole). After stirring for 24 h at rt, the reaction mixture was diluted with brine (20 mL) and extracted with EtOAc (50 mL). The organic layer was washed with 0.1 M NaOH, 0.1 M HCJ, and brine, and dried over MgSO 4 . The organic solvents were removed in vacuo to give 1,6-bis[4-(5-amidopentyloxypropyl) phenoxy]hexane as a pale yellow oily residue (0.45 g). 25 Step 5 A solution of 1,6-bis[4-(5-amidopentyloxypropyl)-phenoxy]hexane (0.45 g, 0.4 mmole) obtained in Step 4 above, in anhydrous THF (10 mL) was added to a solution of LiAlH 4 (0.16 g, 4.22 mmole) in anhydrous THF (40 mL) at 0 0 C . The reaction mixture was stirred for 4 h at 80 'C under nitrogen atmosphere and then 30 quenched by with 10% NaOH (1 mL) at 0*C. After 30 min., the reaction mixture was filtered and the precipitate was washed with 10% MeOH in THF (50 mL). The filtrates were combined and evaporated in vacuo to give a pale yellow oily residue.

WO 01/42193 PCT/USOO/33057 --124- Purification by flash silica column chromatography: 5% MeOH/CH 2 Cl 2 to 3% i PrNH 2 in 10% MeOH/CH 2 Cl 2 gave the 1,6-bis[4-(6-aminohexyloxypropyl) phenoxy]hexane. 'H-NMR (CDCl 3 , 299.96 MHz): 8 (ppm) 7.40-7.25 (in, 12H), 7.22-7.18 (d, 2H), 7.09-7.02 (d, 4H), 6.91-6.88 (d, 2H), 6.81-6.75 (d, 4H), 5.01 (s, 5 4H), 4.8-4.75 (m, 2H), 4.70 (s, 4H), 3.96-3.83 (q, 4H), 3.42-3.34 (in, 8H), 2.84-2.64 (m, 8H), 2.62-2.56 (t, 4H), 1.84-1.75 (in, 8H), 1.57-1.50 (in, 1OH), 1.34-1.23 (m, 1OH). Step 6 A solution of 1,6-bis[4-(6-aminohexyloxypropyl)-phenoxy]hexane (0.16 g, 10 0.15 mmole) obtained in Step 5 above, in EtOH (40 mL) was hydrogenated under

H

2 (1 atm) atmosphere with 10% Pd/C catalyst (100 mg) at rt for 24 h. The catalyst was filtered and the filtrate was concentrated to afford crude product as a pale yellow oil. Purification by reversed phase HPLC: 10 to 50% MeCN/H 2 0 over 40 min; 20 mL/min; 254 nm provides 1,6-bis{4-(N-[2-(4-hydroxy-3-hydroxymethyl 15 phenyl)-2-hydroxyethyl]aminohexyloxypropyl]-phenoxy}hexane 40. H-NMR

(CD

3 0D, 299.96 MHz): 6 (ppm) 7.35 (d, 2H), 7.18-7.15 (dd, 2H), 7.08-7.05 (d, 4H), 6.82-6.77 (m, 6H), 4.65 (s, 4H), 3.96-3.92 (t, 4H), 3.45-3.34 (in, 8H), 3.12 3.01 (in, 6H), 2.94-2.89 (t, 2H), 2.62-2.57 (t, 4H), 1.86-1.43 (in, 28H); ESMS

(C

54

H

80

N

2 0 10 ): calcd. 917.1, obsd. 917.5 [M]*, 940.8 [M+Na]*. 20 Example 6 Synthesis of 1- { 2- [N-2-(4-hydroxy-3 -hydroxymethylphenyl)-2-(R)-hydroxy ethyl]aminoethyl }-4- [N-(2-phenyl-2-(S)-hydroxyethyl)amino]phenyl (following figure 10) 25 OH H N HO HO H O OH Step 1 WO 01/42193 PCT/USOO/33057 --125- A mixture of 4-(N-Boc-2-aminoethyl)aniline 28 (10 g, 42.34 mmole), benzaldehyde (4.52 mL, 44.47 mmole), and molecular sieves 4A (10 g) in toluene (100 mL) was refluxed at 95 "C for 15 h. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give a colorless oil. The oil was dissolved in 5 MeOH (150 mL) and AcOH (0.5 mL) and NaCNBH 3 (2.79 g, 44.4 mmole) were added. The reaction mixture was stirred at 0 C for 1 h and at rt for 2 h and then concentrated in vacuo to give a pale yellow oily residue. Purification by flash silica column chromatography: 1/1 hexane/EtOAc gave N-benzyl-4-(N-Boc-2 aminoethyl)aniline 41 as colorless oil (11.5 g, 83%). Rf = 0.75 in 1/1 10 hexane/EtOAc. H 1 -NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.38-7.2 (in, 5H), 6.87 6.84 (d, 2H), 6.58-6.55 (d, 2H), 4.27 (s, 2H), 3.2-3.15 (in, 2H), 2.6-2.56 (t, 2H), 1.41 (s, 9H); ESMS (C 2 0

H

26

N

2 0 2 ): calcd. 326.4, obsd. 328 [M+H]*. Step 2 A mixture of N-benzyl-4-(N-Boc-2-aminoethyl)aniline 41 (10 g, 30.7 15 mmole) and (R)-styreneoxide (3.51 mL, 30.7 mmole) in EtOH (100 mL) was refluxed for 48 h. A small aliquot of the reaction mixture was taken out for liquid chromatographic analysis, which indicated that the desired adduct 2-[(N-benzyl-4 [2-N-Boc-aminoethyl)anilino] -1 -phenylethanol was formed as a minor product along with another regio-isomer 2-[(N-benzyl-4-[2-N-Boc-aminoethyl)anilino]-2 20 phenyl-ethanol in a ratio of -1/2. Evaporation of the solution afforded thick, pale yellow oil, which was purified by flash silica column chromatography: 4/1 to 2/1 hexane/EtOAc. After repeated chromatography, 2-[(N-benzyl-4-[2-N-Boc aminoethyl)anilino]-1-phenyl-ethanol was obtained as a colorless oil (4.01 g, 29%) (Rf= 0.76 in 2/1 hexane/EtOAc). H'-NMR (CD 3 0D, 299.96 MHz): 6 (ppm) 7.4 25 7.1 (in, 1OH), 7.1-7.06 (d, 2H), 6.68-6.65 (d, 2H), 5.0 (t, 1H), 4.52-4.46 (d, 1H), 4.26-4.22 (d, 1H), 3.76-3.68 (dd, 1H), 3.56-3.48 (dd, 1H), 3.22-3.12 (in, 2H), 2.68 2.56 (in, 2H), 1.41 (s, 9H); ESMS (C 28

H

34

N

2 0 3 ): calcd. 446.6, obsd. 447.1 [M+H]*, 893.4 [2M+H]*. Step 3 30 To a solution of 2-[(N-benzyl-4-[2-N-Boc-aminoethyl)anilino]-1-phenyl ethanol (4.01 g, 8.99 mole) in CH 2 Cl 2 (15 mL) maintained in an ice bath was added TFA (15 mL) under stream of nitrogen atmosphere. After stirring at 0"C for WO 01/42193 PCT/USOO/33057 --126- 30 min., the reaction mixture was concentrated in vacuo, yielding a pale yellow oil. Purification by flash silica column chromatography: (% hexane/EtOAc to 5% i PrNH 2 in 2 hexane/EtOAc) gave 2-[(N-benzyl-4-[2-aminoethyl)anilino]-1-phenyl ethanol 42 as a pale yellow oil from such fractions with Rf of 0.2 (5% i-PrNH 2 in /2 5 hexane/EtOAc) in 74% yield (2.29 g). H'-NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.38-7.06 (in, 10H), 7.01-6.98 (d, 2H), 6.71-6.68 (d, 2H), 5.02-4.96 (dd, 1H), 4.54 4.48 (d, 1H), 4.29-4.23 (d, 1H), 3.76-3.67 (dd, 1H), 3.58-3.50 (dd, 1H), 2.82-2.74 (t, 2H), 2.64-2.59 (t, 2H); ESMS (C 2 3

H

26

N

2 0,): calcd. 346.5, obsd. 346.3[M]*, Step 4 10 A mixture of 2-[(N-benzyl-4-[2-aminoethyl)anilino]-1-phenylethanol 42 (2.28 g, 6.59 mmole), benzaldehyde (0.74 mL, 7.28 mmole), and molecular sieves 4A (4 g) in toluene (40 mL) was heated at 90 'C for 14 h. The reaction mixture was cooled and filtered, and the sieves were rinsed with toluene. The combined filtrates were concentrated to give an oily residue which was washed with hexane, and 15 dried. The residue was dissolved in MeOH (40 mL) containing AcOH (0.4 mL) and the reaction mixture was cooled in an ice bath. NaCNBH 3 (0.62 g, 9.87 mmole) was added and the reaction mixture was stirred for 2 h at rt, and then concentrated. The oily residue was dissolved in 60% MeCN/H 2 0, and purified by reversed phase preparative liquid chromatography (40 to 80% MeCN/H 2 0 over 30 min; 30 20 mL/min) to give 2-[(N-benzyl-4-[2-N-benzylaminoethyl)anilino]-1-phenylethanol as the TFA salt. The product was treated with alkaline brine solution, and extracted with ether (200 mL). The organic layer was dried with NaSO 4 , and concentrated, to give 2-[(N-benzyl-4-[2-N-benzylaminoethyl)anilino]-1 phenylethanol 43 as a colorless oil (1.36 g). H-NMR (CD 3 0D, 299.96 MHz): 6 25 (ppm) 7.36-7.06 (in, 15H), 6.98-6.95 (d, 2H), 6.69-6.60 (d, 2H), 5.01-4.96 (t, 1H), 4.54-4.47 (d, 1H), 4.29-4.24 (d, lH), 3.73 (s, 2H), 3.72-3.68 (dd, IH), 3.59-3.54 (dd, 1H), 2.80-2.74 (in, 2H), 2.70-2.64 (in, 2H); ESMS (C 3 0

H

3 2

N

2 0,): calcd. 436.6, obsd. 437.2 [M+H]*. Step 5 30 A concentrated solution of 2-[(N-benzyl-4-[2-N-benzylaminoethyl)anilino] 1-phenylethanol (1.36 g, 3.12 mmole) and compound (S)-4-benzyloxy-3 methoxycarbonylstyreneoxide 44 (0.887 g, 3.12 mmole; -95% ee) (prepared as WO 01/42193 PCT/USOO/33057 --127described in R. Hett, R. Stare, P. Helquist, Tet. Lett., 35, 9375-9378, (1994)) in toluene (1 mL) was heated at 105 'C for 72 h under nitrogen atmosphere. The reaction mixture was purified by flash silica column chromatography (2/1 hexane/EtOAc to 3% MeOH in 1/1 hexane/EtOAc) to give 1-{2-[N-benzyl-N-2-(4 5 benzyloxy-3-methoxycarbonylphenyl)-2-(R)-hydroxy]ethylaminoethyl}-4-[N-(2 phenyl-2-(S)-hydroxy)ethylamino]benzene 45. (Rf= 0.62 in 3% MeOH in 1/1 hexane/EtOAc) was obtained as a pale yellow foam (2.0 g, 89%). H'-NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.67-7.66 (d, 1H), 7.49-7.42 (m, 2H), 7.38-7.0 (m, 20H), 6.88-6.85 (d, 2H), 6.65-6.62 (d, 2H), 5.15 (s, 2H), 5.05 10 4.98 (t, 1H), 4.6-4.54 (t, 1H), 4.53-4.46 (d, 1H), 4.28-4.22 (d, 1H), 3.84 (s, 3H), 3.72-3.64 (m, 3H), 3.56-3.46 (dd, 1H), 2.74-2.56 (m, 6H); ESMS (C 47

H

4 8

N

2 0 5 ): calcd. 720.9, obsd. 721.4 [M+H]*, 743.3 [M+Na]*. Step 6 To a suspension of LiAlH 4 (0.211 g, 5.56 mmole) in THF (40 mL) cooled 15 with ice bath was added 1-{2-[N-benzyl-N-2-(4-benzyloxy-3 methoxycarbonylphenyl)-2-(R)-hydroxyethyl]aminoethyl}-4-[N-(2-phenyl-2-(S) hydroxyethyl)amino]benzene 45 (2.0 g, 2.78 mmole) in THF (10 mL) under nitrogen atmosphere. The reaction mixture was warmed slowly to rt and the stirring was continued for 5 h. The reaction was cooled to 0 C, and 10% NaOH (0.5 mL) 20 was slowly added. After 30 min., a thick gel formed. The gel was diluted with THF (300 mL), filtered, and the solid mass was rinsed with THF (50 mL). The filtrates were combined, and concentrated in vacuo, yielding an oily residue. The residue was purified by flash silica column chromatography (2/1 hexane/EtOAc to 3% MeOH in 1/1 hexane/EtOAc) to give 1-{2-[N-benzyl-N-2-(4-benzyloxy-3 25 hydroxymethylphenyl)-2-(R)-hydroxyethyl]aminoethyl } -4- [N-(2-phenyl-2-(S) hydroxyethyl)amino]benzene as a colorless oil (1.28 g, 67%). H 1 -NMR (CD 3 0D, 299.96 MHz): 6 (ppm) 7.4-7.0 (m, 22H), 6.85-6.82 (m, 3H), 6.63-6.60 (d, 2H), 5.02-4.94 (m, 3H), 4.66 (s, 2H), 4.59-4.54 (dd, 1H), 4.48-4.4 (d, 1H), 4.24-4.16 (d, 1H), 3.76-3.7 (d, 1H), 3.69-3.62 (dd, 1H), 3.58-3.52 (d, 1H), 3.50-3.44 (dd, 1H), 30 2.76-2.54 (m, 6H); ESMS (C 4 6

H

4 8

N

2 0 4 ): calcd. 692.90, obsd. 693.5 [M+H]*. Step 7 WO 01/42193 PCT/USOO/33057 --128- A solution of 1-{2-[N-benzyl-N-2-(4-benzyloxy-3-hydroxymethylphenyl)-2 (R)-hydroxyethyl]amino] ethyl} -4- [N-(2-phenyl-2-(S)-hydroxyethyl)amino] -benzene (1.28 g, 1.85 mmole) in EtOH (80 mL) was hydrogenated under H 2 (1 atm) with 10% Pd/C (0.6 g) for 36 h. After filtration and rinsing of the catalyst with EtOH 5 (50 mL), the filtrates were combined, and evaporated in vacuo, yielding pale yellow foam which was dissolved in 10% MeCN/H 2 0, and purified by reversed phase preparative liquid chromatography (10 to 30% MeCN/H 2 0 (containing 0.3% TFA) over 50 min; 30 mL/min; 254 nm) to give 1-{2-[N-2-(4-hydroxy-3-hydroxymethyl phenyl)-2-(R)-hydroxyethyl]aminoethyl}-4-[N-(2-phenyl-2-(S)-hydroxyethyl) 10 amino]benzene as the TFA salt (0.6 g, 50%). Optical purity of 1-{2-[N-2-(4 hydroxy-3-hydroxymethylphenyl)-2-(R)-hydroxyethyl]aminoethyl}-4-[N-(2-phenyl 2-(S)-hydroxyethyl)amino]benzene 46 which was analyzed with capillary electrophoresis by using a chiral medium, and estimated to be ~93%. H'-NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.42-7.28 (m, 8H), 7.26-7.22 (d, 15 2H), 7.18-7.14 (dd, 1H), 6.80-6.77 (d, 1H), 4.88-4.82 (m, 2H), 4.65 (s, 2H), 3.5 3.43 (m, 2H), 3.29-3.26 (in, 2H), 3.19-4.14 (m, 2H), 3.06-3.0 (m, 2H); ESMS

(C

2 5

H

30

N

2 0 4 ): calcd. 422.5, obsd. 423.1 [M+H]*, 445.4 [M+Na]*. Example 7 20 Synthesis of 1-{6-[N-[2-(4-hydroxy-3-hydroxymethylphenyl)-2-[hydroxyethyl] amino]hexyloxy}-4-{6-[N-[2-(4-hydroxy-3-hydroxy-methylphenyl)-2 hydroxyethyl] amino]hexyloxypropyl} benzene (following figure 11) H HO OH HON OHH O OH HO HO HO Step 1 25 A solution of 3-(4-hydroxyphenyl)- 1 -propanol (2.0 g, 13.1 mmole) in DMF (5 mL ) was added to a solution of DMF (35 mL) containing NaH (1.31 g, 60% in mineral oil) at 0"C under nitrogen atmosphere. The reaction mixture was slowly WO 01/42193 PCT/USOO/33057 --129warmed to 80 *C. After stirring for 1 h at 80 "C, the reaction mixture was cooled to 0 'C, and 6-bromohexanenitrile (5.78 g, 32.83 mmole) was added. The final mixture was re-heated to 80 "C, and stirred for 24 h. The reaction mixture was quenched with saturated NaCl solution (200 mL), and the product was extracted 5 with EtOAc (300 mL). The organic layer was washed with brine solution, dried with Na2SO4, and evaporated to dryness, yielding a pale yellow solid. Purification of the crude product by flash silica column chromatography: 4/1 to 1/1 hexane/EtOAc provided 6-{3-[4-(5-cyanopentyloxy)phenyl]propoxy}hexanenitrile in 30% yield (1.33 g). Rf= 0.63 in 1/1 EtOAc/hexane. 'H-NMR (CDCl 3 , 299.96 10 MHz): 5 (ppm) 7.09-7.07 (d, 2H), 6.81-6.78 (d, 2H), 3.96-3.92 (t, 2H), 3.42-3.37 (in, 4H), 2.64-2.58 (t, 2H), 2.39-2.32 (in, 4H), 1.87-1.52 (in, 14 H). Step 2 A solution of 6-{3-[4-(5-pentyloxy)phenyl]propoxy}hexanenitrile (1.33 g, 3.88 mmole) in THF (10 mL) was added to a solution of LiAlH 4 (0.442 g, 11.65 15 mmole) in THF (50 mL) at 0 *C under nitrogen atmosphere. The reaction mixture was heated slowly to reflux, and stirred for 2 h. The reaction mixture was cooled to 0 *C, and 10% NaOH solution (5 mL) was slowly added. After 30 min., the reaction mixture was filtered, and the collected solids were washed with THF (100 mL). The filtrate was concentrated to yield a pale yellow oil which was purified by 20 flash silica column chromatography: 5% MeOH/CH 2 Cl2 to 3% i-PrNH 2 /20% MeOH/CH 2 Cl 2 to give 6-{3-[4-(6-aminohexyloxy)-phenyl]propoxy}-hexylamine as a colorless oil (0.5 g, 37%) which was converted to the desired compound by proceeding as described in Example 1, step 2 above. The crude product was purified by preparatory reversed phase HPLC: 10 to 40% MeCN/H 2 0 over 40 min; 25 20 mL/min; 254 nm. ESMS (C 39

H

58

N

2 0 8 ): calcd. 682.8, obsd. 683.6 [M+H]*, 797.5

[M+CF

3

CO

2 H]*. Example 8 Synthesis of bis{2-{2-(4-hydroxy-3-hydroxymethylphenyl)-2-hydroxy]ethyamino} 30 2-hydroxyethoxy} benzene (following figure 12) WO 01/42193 PCT/USOO/33057 --130- OH OH HN 0 HO OH HO H 5 HO 0 N OH OH Step 1 To a N 2 -saturated solution of acetonitrile (300 mL) containing methyl 5 10 acetylsalicylate 50 (20 g, 0.1 mole) and benzylbromide (13.5 mL, 0.11 mole) was added K 2

CO

3 (28.5 g, 0.21 mole). The reaction mixture was stirred at 90 0 C for 5 h. After cooling, the reaction mixture was filtered, and the filtrate was concentrated, in vacuo, yielding a white solid which was susended in hexane (300 mL), and collected on Buchner funnel to give methyl 0-benzyl-5-acetylsalicylate 51 as 15 colorless to white crystals (28.1 g, 96%). Rf= 0.69 in 1/1 EtOAc/hexane. H'-NMR (CDCl 3 , 299.96 MHz): 8 (ppm) 7.8.43-8.42 (d, 1H), 8.1-8.04 (dd, 1H), 7.5-7.28 (m, 5H), 7.08-7.04 (d, 1H), 5.27 (s, 2H), 3.93 (s, 3H), 2.58 (s, 3H). Step 2 To a solution of methyl 0-benzyl-5-acetylsalicylate 51 (14.15 g, 0.05 mole) 20 in CHCl 3 (750 mL) was added bromine (2.70 mL, 0.052 mole). The reaction mixture was stirred at rt. While being stirred, the reaction mixture gradually turned from red-brown to colorless. The mixture was stirred for 2 h at rt, and quenched by adding brine solution (300mL). After shaking the mixture in a separatory funnel, organic layer was collected, washed with brine, and dried under Na 2

SO

4 . The 25 organic solution was concentrated in vacuo, yielding white solid. It was washed with ether (200 mL). After drying in air, 15 g (83%) of methyl 0-benzyl-5 (bromoacetyl)-salicylate 52 was obtained. Rf = 0.76 in 1/1 EtOAc/hexane. H' NMR (CDCl 3 , 299.96 MHz): S (ppm) 8.48-8.46 (d, 1H), 8.14-8.08 (dd, 1H), 7.51 7.3 (m, 5H), 7.12-7.09 (d, 1H), 5.29 (s, 2H), 4.42 (s, 2H), 3.94 (s, 3H). 30 Step 3 To a solution of DMF (60 mL) containing methyl 0-benzyl-5 (bromoacetyl)-salicylate 52 (7.08 g, 0.019 mole) was added NaN 3 (1.9 g, 0.029 WO 01/42193 PCT/USOO/33057 --13 1mole). After stirring at rt for 24 h in the dark, the mixture was diluted with EtOAc (200 mL), and washed with brine solution (3 x 200 mL) in a separatory funnel. The organic phase was dried under MgSO 4 , and concentrated to afford pale red solid. It was purified by flash silica column chromatography: 10 to 50% EtOAc in hexane. 5 The desired product methyl 0-benzyl-5-(azidoacetyl)salicylate 53 was obtained as white crystals (4.7 g, 74%). Rf= 0.68 in 1/1 EtOAc/hexane. H'-NMR (CDCl 3 , 299.96 MHz): 8 (ppm) 8.38-8.36 (d, 1H), 8.08-8.04 (dd, 1H), 7.5-7.3 (in, 5H), 7.12 7.09 (d, 1H), 5.29 (s, 2H), 4.53 (s, 2H), 3.94 (s, 3H). Step 4 10 To a gray suspension of LiAlH 4 (2.74 g, 0.072 mole) in THF (400 mL) cooled in ice bath was added methyl 0-benzyl-5-(azidoacetyl)salicylate 53 (4.7 g, 0.014 mole) under nitrogen atmosphere. The reaction mixture was stirred at 0 0 c for 1 h, and gradually warmed to rt. After stirring for 16 h at rt, the mixture was heated at 751c for 3 h. The reaction mixture was cooled in ice bath, and quenched by 15 slowly adding 10% NaOH (10 mL). After stirring for 1 h, precipitates were filtered, and rinsed with 5% MeOH in THF (200 mL). Filtrates were combined, and concentrated in vacuo, yielding pale yellow oily residue. The crude product was purified by flash silica column chromatography: 10% MeOH/CH 2 Cl 2 to 5% i PrNH 2 in 30% MeOH/CH 2 Cl 2 to give 2-(4-benzyloxy-3-hydroxymethylphenyl)-2 20 hydroxyethylamine 39 as a pale yellow solid (2.6 g, 66%). Rf = 0.63 in 5% i-PrNH2 in 30% MeOH/CH 2 Cl 2 . H-NMR (CD 3 0D, 299.96 MHz): 8 (ppm) 7.46-7.28 (in, 6H), 7.24-7.20 (dd, 1H), 7.0-6.96 (d, 1H), 5.11 (s, 2H), 4.70 (s, 2H), 4.65-4.60 (t, 1H), 2.83-2.81 (d, 2H); ESMS (C 1

H

19 Ni0 3 ): calcd. 273.3, obsd. 274.7 [M+H]*, 547.3 [2M+H]*. 25 Step 5 To a solution of EtOH (15 mL) containing compound 2-(4-benzyloxy-3 hydroxymethylphenyl)-2-hydroxyethylamine 39 (0.3 g, 1.1 mmole) was added resorcinol diglycidyl ether (0.122 g, 0.55 mmole) dissolved in EtOH (5 mL). The reaction mixture was refluxed for 20 h. After cooling down to rt, the reaction 30 mixture was degassed with nitrogen and hydrogenated with 10% Pd/C (0.3 g, 10%) under H 2 (1 atm) atmosphere for 24 h. After filtration of the catalyst, the filtrate was concentrated to dryness, yielding a colorless oily residue which was purified by WO 01/42193 PCT/USOO/33057 --132preparatory reversed phase HPLC (10 to 50% MeCN/H 2 0 over 40 min; 20 mL/min; 254 nm) to give bis{2-{2-(4-hydroxy-3-hydroxymethylphenyl)-2-hydroxy] ethyamino} -2-hydroxyethoxy} benzene 54. ESMS (C 30

H

4 0

N

2 0 1 o): calcd. 588.6, obsd. 589.4 [M+H]*, 610.7 [M+Na]*. 5 Example 9 Synthesis of 1-{2-[N-2-(4-hydroxy-3-hydroxymethylphenyl)-2-hydroxy-ethyl] amino]ethyl} -4- [N-(2-napth- 1 -yloxymethyl-2-hydroxyethyl)amino]benzene (following figure 13) OH H N HON HO OH 10 Step 1 A solution of EtOH (50 mL) containing 4-(N-Boc-2-aminoethyl)aniline 28 (0.4 g, 1.69 mmole) and 3-(1-naphthoxy)-1,2-epoxypropane 55 (0.33 g, 1.65 mmole) was refluxed for 18 h, and concentrated in vacuo to dryness, yielding a pale yellow oil. It was dissolved in 10 mL of CH 2 Cl 2 , cooled in ice bath, and treated with TFA 15 (5 mL). After stirring for 2 h at 0 0 C, the mixture was evaporated, yielding a pale red oil. It was dissolved in 30% aqueous acetonitrile, and purified by preparatory HPLC: 10 to 30% MeCN/H 2 0 over 30 min; 20 mL/min; 254 nm. The product 56 was obtained as colorless oil (260 mg; TFA salt). H'-NMR (CD 3 0D, 299.96 MHz): d (ppm) 8.88-8.25 (dd, 1H), 7.82-7.79 (dd, 1H), 7.51-7.42 (in, 3H), 7.39 20 7.38 (d, 1H), 7.33-7.30 (d, 2H), 7.25-7.23 (d, 2H), 6.91-6.89 (d, 1H), 4.37-4.31 (m, 1H), 4.22-4.19 (in, 2H), 3.69-3.63 (dd, 1H), 3.67-3.54 (dd, 1H), 3.17-3.11 (t, 2H), 2.96-2.91 (t, 2H); ESMS (C 2 1

H

24

N

2 0 2 ): calcd. 336.4, obsd. 337.5 [M+H]*, 359.6 [M+Na]*, 673.4 [2M+H]*. Step 2 25 To a solution of compound 56 (0.13 g, 0.023 mmole; TFA salt) in 5 mL of MeOH was added 1.0 M NaOH (1.0 M, 0.46 mL). After homogeneous mixing, the solution was evaporated to dryness. The residue was dissolved in THF (10 mL), WO 01/42193 PCT/USOO/33057 --133followed by addition of glyoxal 12 (52 mg; 0.023 mmole). The resulting suspension was stirred for 4 h at ambient temperature under nitrogen atmosphere. After cooling of the resulting solution in ice bath, an excess amount of 2M BH 3 Me 2 S in THF (3 mL; 6 mmole) was added to the previous reaction solution. The 5 resulting mixture was slowly warmed to rt, and refluxed for 4 h under N 2 stream. After cooling of the hot solution, 5 mL of MeOH was added to the cooled mixture to quench the reaction mixture under nitrogen atmosphere. After stirring 30 min at rt, the final solution was evaporated in vacuo, yielding a pale brown solid. It was washed with EtOAc/hexane (1/2; 20 mL), and dried. The 10 crude product was dissolved in 50% MeCN/H 2 0 containing 0.5% TFA, and purified by prep-scale high performance liquid chromatography (HPLC) using a linear gradient (5% to 50% MeCN/H 2 0 over 50 min, 20 mL/min; detection at 254 nM). Fractions with UV absorption were analyzed by LC-MS to locate the desired product 1-{2-[N-2-(4-hydroxy-3-hydroxy-methylphenyl)-2-hydroxyethyl]amino] 15 ethyl} -4- [N-(2-napth- 1 -yloxymethyl-2-hydroxy-ethyl)amino]benzene 57. ESMS

(C

30

H

3 4

N

2 0 5 ): calcd. 502.6, obsd. 503.2 [M+H]*, 525.6 [M+Na]*. Example 10 20 Synthesis of 1,4,7-tris{N-[2-(4-hydroxy-3-hydroxymethylphenyl)-2 hydroxyethyl]amino} octane OH OH N H OH OH OH 9" ' "' OH H O HO To a suspension of a,a-dihydroxy-4-hydroxy-3-methoxycarbonyl acetophenone 12 (0.45 g, 1.99 mmol) in tetrahydrofuran (15 mL) was added a 25 solution of 4-(aminomethyl)-1,8-octadiamine (115 mg, 0.66 mmol) in WO 01/42193 PCT/USOO/33057 --134tetrahydrofuran (5 mL). The resulting suspension was stirred for 12 h at ambient temperature under nitrogen atmosphere. After cooling of the resulting solution in ice bath an excess amount of 2 M BH 3 -Me 2 S in hexane (6 mL, 12 mmol) was added. The resulting mixture was slowly warmed to rt, and refluxed for 6 h under 5 nitrogen atmosphere. After cooling, the reaction mixture was quenched with methanol (5 mL). The resulting solution was stirred at rt for 30 min., and then concentrated in vacuo to give a pale brown solid. The solid was washed with ethyl acetate :hexane mixture (1:2) and then dried. The crude product was dissolved in 50% acetonitrile/water containing 0.5% TFA and purified by HPLC using a linear 10 gradient (5% to 50% MeCN/H 2 0 over 50 min., 20 mL/min.; detection at 254 nM). Fractions with UV absorption was analyzed by LC-MS to locate the desired product. ESMS (C 36

H

5 3

N

3 0 9 ): Calcd. 671.8; Obsd. 671.7. Example 11 15 Synthesis of 1-{ 2-[N-2-(4-amino-3,5-dichlorophenyl)-2-(RS) hydroxyethyl]aminoethyl}-4-[N-(2-phenyl-2-(RS)-hydroxyethyl)amino]phenyl (following figure 14) OH H 20 CI N C I 0H Step 1 25 A solution of 2-(4-aminophenyl)ethylamine 25 (4.70 mL, 36.7 mmol), benzaldehyde (7.46 mL, 73.4 mmol) and 4A molecular sieves (18 g) in toluene (180 mL) was refluxed for 3 h. The reaction mixture was cooled and filtered. The filtrate was concentrated under reduced pressure to afford compound 58 (95% yield). 30 Step 2 To a cooled a solution of 58 (2.00 g, 6.40 mmol) in ethanol (150 mL) in ice bath was slowly added sodium borohydride (361 mg, 9.50 mmol) under a nitrogen WO 01/42193 PCT/USOO/33057 --135atmosphere. The reaction mixture was allowed to stir at 0 *C for 1.5 h, and then warmed slowly to room temperature. The reaction mixture was quenched by slowly adding 50% methanol/TFA (5 mL) and then the mixture was concentrated under reduced pressure. The resultant residue was dissolved in EtOAc, and 5 washed with 0.1 M NaOH. After drying over Na 2

SO

4 , the organic phase was concentrated in vacuo, and the residue was purified by flash silica gel chromatography using ethyl acetate/hexanes as eluant to give compound 59(80% yield). Step 3 10 To a solution of 59 (1.00 g, 3.20 mmol) in methanol cooled with ice bath was slowly added di-tert-butyl dicarbonate (0.69 g, 3.2 mmol) in cold methanol (5 mL). The reaction mixture was allowed to stir at 00 C for 0.5 h, and then warmed gradually to room temperature. After stirring for 1 h, the reaction mixture was concentrated under reduced pressure, and dried under high vacuum overnight. The 15 oily residue was purified by silica gel chromatography using ethyl acetate/ hexane (2:1) to afford compound 60 (80% yield). Step 4 To a solution of 60 (2.09 g, 5.00 mmol) in methanol (45 mL) was added phenylglyoxal (2.01 g, 15.0 mmol). The reaction mixture was stirred at room 20 temperature for I h, and followed by slow addition of sodium cyanoborohydride (1.25 g, 20 mmol). The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in methanol. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography 25 using hexane/ ethyl acetate (9:1) to afford compound 61 (26% yield). Step 5 To a cooled a solution of 61 (3.19 g, 5.90 mmol) in methylene chloride (12 mL) in an ice bath was added slowly trifluoroacetic acid (12 mL) under stream of nitrogen. After stirring the reaction mixture for 1 h at the same temperature, the 30 mixture was concentrated under reduced pressure to yield compound 62 as an oily residue. The product was dried in vacuo overnight, and was used in next step without further purification (90% yield).

WO 01/42193 PCT/USOO/33057 --136- Step 6 To a mixture of 62 (1.87 g, 3.4 mmol) and 2,6-dichloro-4-(bromoacetyl) aniline 63 (1.07 g, 3.8 mmol) in DMF (50 mL) was added potassium carbonate (0.96 g, 6.90 mmol). The reaction mixture was stirred at room temperature for 0.5 5 h and then at 35 "C for 1 h. The reaction mixture was cooled to room temperature, and sodium borohydride (0.16 g, 4.10 mmol) in ethanol (50mL) was added slowly. The reaction continued overnight at room temperature and then quenched with aqueous NH 4 C1 (sat'd). After concentration of the reaction mixture under reduced pressure, the residue was dissolved in ether, and washed with brine. After drying 10 over MgSO 4 , the organic layer was concentrated, and the residue was purified by silica gel chromatography by eluting with hexane/ethyl acetate (4:1). The product 64 was obtained in 50% yield. Step 7 A suspension of 64 (107 mg, 0.17 mmol) and palladium hydroxide (25 mg) 15 in ethanol (2.5 mL) was stirred overnight under hydrogen atmosphere (1 atm) at room temperature. The mixture was filtered, and the filtrate was concentrated to yield crude product which was purified using silica gel chromatography with 6% methanol/dichloromethane (10% yield) to give compound 65. ESMS

(C

24

H

27 Cl 2

N

3 0 2 ): calcd: 460.4; obsd: 460 [M+H]*, 442 [M-H 2 0]*, 921 [2M+H] *. 20 Example 12 Synthesis of 1-{2-[N-2-(4-hydroxy-3-formylaminophenyl)-2-(RS) hydroxyethyl]aminoethyl}-4-[N-(2-phenyl-2-(RS)-hydroxyethyl)amino]phenyl 25 (following figure 15) OH H N HO N N NHCHO H OH WO 01/42193 PCT/USOO/33057 --137- Step 1 1-Benzyloxy-4-bromoacetyl-2-nitrobenzene 66 (3.76 g, 10.8 mmol) [prepared as described in Chem. Bull., 25, 1368-1377, (1977)], was added to a solution of compound 59 (3.4 g, 10.8 mmol) in dimethylformamide (150 mL) at 5 room temperature. After 28 h, the reaction mixture was diluted with ether and washed with a dilute solution of aqueous sodium chloride. The organic layer was separated and dried over sodium sulfate, filtered and concentrated to give a crude oil. Purification with column chromatography using hexane:ethyl acetate as the eluent provided compound 67 (95% yield). 10 Step 2 To a solution of 67 (4.0 g, 6.80 mmol) in ethanol (500 mL) was slowly added sodium borohydride (1 g, 26.50 mmol) in portions over 30 min., under a nitrogen atmosphere. After 6 h, the reaction mixture was quenched by slowly adding saturated solution of aqueous ammonium chloride . The solution was diluted with 15 1 N sodium hydroxide and ethyl acetate and hexanes. The organic layer was separated, dried over sodium sulfate, filtered and concentrated to give compound 68 as an oil which was used in the next step without further purification. Step 3 To a mixture of 68 (3.6 g, 6.1 mmol) and potassium carbonate (3.0 g, 9.2 20 mmol) in dimethylformamide (100 mL) was added alpha-bromoacetophenone (1.28 g, 6.4 mmol) portionwise. The reaction mixture was heated overnight at 65 *C. An additional portion of alpha-bromoacetophenone (0.25 g, 1.25 mmol) was added and heating was continued. After 18 h, the reaction mixture was cooled to room temperature and ethanol (50 mL) was added. Sodium borohydride (1.0 g, 25 26.5 mmol) was added and stirring was continued for 2.5 h. The reaction mixture was concentrated and then methanol (25 mL) was added and the excess hydride was quenched with the addition of a saturated solution of ammonium chloride. The reaction mixture was diluted with ethyl acetate and ether. The organic layer was separated, dried over sodium sulfate, filtered, and concentrated. The crude oil 30 was purified with column chromatography eluting with ethyl acetate/hexanes mixture to give compound 69. Step 4 WO 01/42193 PCT/USOO/33057 --138- To a solution of 69 (0.73 g, 1.0 mmol) in a mixture of methanol (20 mL), 6 N hydrochloride acid (1 mL) and water (2 mL) was added iron powder (0.56 g, 10.0 mmol). The reaction mixture was heated at 90 *C for 1.5 h. The reaction mixture was cooled to room temperature and allowed to stand overnight. Methanol was 5 added and the brown precipitates and unreacted iron was filtered off. The filtrate was concentrated under reduced pressure, to give 70 as a brown solid which was used in the next step without further purification. Step 5 Compound 70 was dissolved in a premixed solution of acetic anhydride (5 10 mL) and formic acid (3 mL) at room temperature. After 3 h, the reaction mixture was diluted with ethyl acetate and evaporated to dryness. A methanolic solution of 0.5 M sodium hydroxide was added and the reaction mixture was stirred for 6 h at room temperature. The reaction mixture was treated with methanolic solution of 1 N hydrochloric acid and then concentrated to dryness. The residue was redissolved 15 in methanol and filtered. The filtrate was concentrated to give compound 71 as a brown residue which was used in the next step without further purification. Step 6 Palladium on carbon (10%, 0.5 g) was added to a suspension of 71 in methanol (120 mL) and dimethylformamide (80 mL). The reaction mixture was 20 purged with nitrogen gas and stirred overnight under hydrogen atmosphere (1 atm) at room temperature. The mixture was filtered, and the filtrate was concentrated to yield crude product which was purified by HPLC (acetonitril/water/1% TFA gradient) to give crude product which was purified with column chromatography using methanol/methylene chloride/1%isopropylamine as the eluent to give desired 25 compound 1-{2-[N-2-(4-hydroxy-3-formylaminophenyl)-2-(RS)-hydroxyethyl] aminoethyl}-4-[N-(2-phenyl-2-(RS)-hydroxyethyl)amino]phenyl 72. Formulation Examples Example 1 30 Hard gelatin capsules containing the following ingredients are prepared: Quantity Ingredient (mg/capsule) WO 01/42193 PCT/USOO/33057 --139- Active Ingredient 30.0 Starch 305.0 Magnesium stearate 5.0 5 The above ingredients are mixed and filled into hard gelatin capsules in 340 mg quantities. Example 2 A tablet Formula is prepared using the ingredients below: 10 Quantity Ingredient (mg/tablet) Active Ingredient 25.0 Cellulose, microcrystalline 200.0 15 Colloidal silicon dioxide 10.0 Stearic acid 5.0 The components are blended and compressed to form tablets, each weighing 240 mg. 20 Example 3 A dry powder inhaler formulation is prepared containing the following components: Ingredient Weight % 25 Active Ingredient 5 Lactose 95 The active ingredient is mixed with the lactose and the mixture is added to 30 a dry powder inhaling appliance. Example 4 Tablets, each containing 30 mg of active ingredient, are prepared as follows: 35 Quantity Ingredient (mg/tablet) Active Ingredient 30.0 mg WO 01/42193 PCT/USOO/33057 --140- Starch 45.0 mg Microcrystalline cellulose 35.0 mg Polyvinylpyrrolidone (as 10% solution in sterile water) 4.0 mg 5 Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1.0 mg Total 120.0 mg 10 The active ingredient, starch and cellulose are passed through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders, which are then passed through a 16 mesh U.S. sieve. The granules so produced are dried at 500 to 60'C and passed through a 16 15 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 30 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 120 mg. 20 Example 5 Capsules, each containing 40 mg of medicament are made as follows: Quantity Ingredient (mg/capsule) Active Ingredient 40.0 mg 25 Starch 109.0 mg Magnesium stearate 1.0 mg Total 150.0 mg The active ingredient, starch, and magnesium stearate are blended, passed 30 through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 150 mg quantities. Example 6 35 Suppositories, each containing 25 mg of active ingredient are made as follows: Ingredient Amount Active Ingredient 25 mg Saturated fatty acid glycerides to 2,000 mg WO 01/42193 PCT/USOO/33057 --141- The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2.0 g capacity and allowed to cool. 5 Example 7 Suspensions, each containing 50 mg of medicament per 5.0 mL dose are made as follows: Ingredient Amount 10 Active Ingredient 50.0 mg Xanthan gum 4.0 mg Sodium carboxymethyl cellulose (11%) Microcrystalline cellulose (89%) 50.0 mg Sucrose 1.75 g 15 Sodium benzoate 10.0 mg Flavor and Color q.v. Purified water to 5.0 mL The active ingredient, sucrose and xanthan gum are blended, passed through 20 a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of the microcrystalline cellulose and sodium carboxymethyl cellulose in water. The sodium benzoate, flavor, and color are diluted with some of the water and added with stirring. Sufficient water is then added to produce the required volume. 25 Example 8 A formulation may be prepared as follows: Quantity Ingredient (mg/capsule) 30 Active Ingredient 15.0 mg Starch 407.0 mg Magnesium stearate 3.0 mg Total 425.0 mg 35 The active ingredient, starch, and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 425.0 mg quantities.

WO 01/42193 PCT/USOO/33057 --142- Example 9 A formulation may be prepared as follows: Ingredient Ouantity Active Ingredient 5.0 mg 5 Corn Oil 1.0 mL Example 10 A topical formulation may be prepared as follows: 10 Ingredient Ouantity Active Ingredient 1-10 g Emulsifying Wax 30 g Liquid Paraffin 20 g White Soft Paraffin to 100 g 15 The white soft paraffin is heated until molten. The liquid paraffin and emulsifying wax are incorporated and stirred until dissolved. The active ingredient is added and stirring is continued until dispersed. The mixture is then cooled until solid. 20 Another preferred formulation employed in the methods of the present invention employs transdermal delivery devices ("patches"). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds of the present invention in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known 25 in the art. See, e.g., U.S. Patent 5,023,252, issued June 11, 1991, herein incorporated by reference in its entirety. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents. Other suitable formulations for use in the present invention can be found in Remington's Pharmaceutical Sciences, edited by E. W. Martin (Mack Publishing 30 Company, 18th ed., 1990). Biological Examples Example 1 02-Adrenergic Receptor In Vitro Functional Assay 35 The P2-adrenergic receptor functional activity of compounds of the WO 01/42193 PCT/USOO/33057 --143invention was tested follows. Cell Seeding and Growth: Primary bronchial smooth muscle cells from a 21 yr. old male (Clonetics, San Diego, CA) were seeded at 50,000 cells/well in 24-well tissue 5 culture plates. The media used was Clonetic's SmBM-2 supplemented with hEGF, Insulin, hFGF, and Fetal Bovine Serum. Cells were grown two days at 37*C, 5% CO 2 until confluent monolayers were seen. Agonist Stimulation of Cells The media was aspirated from each well and replaced with 250 ml fresh media 10 containing 1mM IBMX, a phospodiesterase inhibitor (Sigma, St Louis, MO). Cells were incubated for 15 minutes at 37 *C, and then 250 ml of agonist at appropriate concentration was added. Cells were then incubated for an additional 10 minutes. Media was aspirated and 500 ml cold 70% EtOH was added to cells, and then removed to an empty 96-well deep-well plate after about 5 minutes. This 15 step was then repeated. The deep-well plate was then spun in a speed-vac until all EtOH dried off, leaving dry pellets. cAMP (pmol/well) was quantitated using a cAMP ELISA kit from Stratagene (La Jolla, CA). EC 50 curves were generated using the 4-parameter fit equation: y = (a-d)/(1+ (x/c)) + d, where, 20 y = cpm a = total binding c = IC 50 x = [compound] d = NS binding b = slope Fix NS binding and allow all other parameters to float. 25 Example 2 P2-Adrenergic Receptor In Vitro Radioligand Binding Assay The 1/2-adrenergic receptor binding activity of compounds of the invention can be tested follows. SF9 cell membranes containing either P l or p2-adrenergic receptor (NEN, Boston, MA) were incubated with 0.07 nM 30 '"-iodocyanopindolol (NEN, Boston, MA) in binding buffer containing 75mM Tris-HCl (pH 7.4), 12.5 mM MgCl 2 and 2 mM EDTA and varying concentrations of test compounds or buffer only (control) in 96-well plates. The plates were WO 01/42193 PCT/USOO/33057 --144incubated at room temperature with shaking for 1 hour. The receptor bound radioligand was harvested by filtration over 96-well GF/B filter plates (Packard, Meriden, CT) pre-blocked with 0.3%polyethylenimine and washed twice with 200pl PBS using cell harvester. The filters were washed thrice with 200pl PBS 5 using cell harvester and then resuspended in 4 0pl scintillation cocktail. The filter bound radioactivity was measured with a scintillation counter and IC 50 curves are generated using the standard 4-parameter fit equation described above. The foregoing invention has been described in some detail by way of 10 illustration and example, for purposes of clarity and understanding. It will be obvious to one of skill in the art that changes and modifications may be practiced within the scope of the appended claims. Therefore, it is to be understood that the above description is intended to be illustrative and not restrictive. The scope of the invention should, therefore, be determined not with reference to the above 15 description, but should instead be determined with reference to the following appended claims, along with the full scope of equivalents to which such claims are entitled.

Claims (16)

1. A compound of the formula: 5 OH H Arl H OH 10 wherein: Ar' is selected from the group consisting of 4-amino-3,5-dichlorophenyl and

3-formylamino-4-hydroxyphenyl; and the stereochemistry at *C and **C is (RS) and (RS); (R) and (R); (S) and (S); (R) and (S); or (S) and (R); and pharmaceutically acceptable salts thereof. 15 2. The compound of Claim 1, wherein Ar is 4-amino-3,5-dichloro phenyl, and the stereochemistry at *C is (RS) and the stereochemistry at **C is (RS). 20 3. The compound of Claim 1, wherein Ar' is 4-amino-3,5-dichloro phenyl, and the stereochemistry at *C is (R) and the stereochemistry at **C is (R).

4. The compound of Claim 1, wherein Ar' is 4-amino-3,5-dichloro phenyl, and the stereochemistry at *C is (S) and the stereochemistry at **C is (S). 25

5. The compound of Claim 1, wherein Ar' is 4-amino-3,5-dichloro phenyl, and the stereochemistry at *C is (R) and the stereochemistry at **C is (S).

6. The compound of Claim 1, wherein Ar' is 4-amino-3,5-dichloro 30 phenyl, and the stereochemistry at *C is (S) and the stereochemistry at **C is (R).

7. The compound of Claim 1, wherein Ar is 3-formylamino-4- WO 01/42193 PCT/USOO/33057 --146- hydroxy-phenyl, and the stereochemistry at *C is (RS) and the stereochemistry at C is (RS).

8. The compound of Claim 1, wherein Ar' is 3-formylamino-4 5 hydroxy-phenyl, and the stereochemistry at *C is (R) and the stereochemistry at **C is (R).

9. The compound of Claim 1, wherein Ar' is 3-formylamino-4 hydroxy-phenyl, and the stereochemistry at *C is (S) and the stereochemistry at 10 **C is (S).

10. The compound of Claim 1, wherein Ar' is 3-formylamino-4 hydroxy-phenyl, and the stereochemistry at *C is (R) and the stereochemistry at **C is (S). 15

11. The compound of Claim 1, wherein Ar' is 3-formylamino-4 hydroxy-phenyl, and the stereochemistry at *C is (S) and the stereochemistry at **C is (R). 20 12. A compound of the formula: OH H CI N H N N 25 2 H CI OH wherein the stereochemistry at *C and **C is RS, R or S; and pharmaceutically acceptable salts thereof. 30

13. A compound of the formula: WO 01/42193 PCT/USOO/33057 --147- OH H N HO N H 5 NHCHO OH wherein the stereochemistry at *C and **C is RS, R or S; and pharmaceutically acceptable salts thereof. 10 14 A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of any of Claims 1-13.

15. The pharmaceutical composition of Claim 14, wherein the 15 pharmaceutical composition further comprises a therapeutically effective amount of a steroidal anti-inflammatory drug.

16. A method for treating diseases mediated by a p2 adrenergic receptor in a mammal, the method comprising administering to the mammal a 20 pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of any of Claims 1-13.

17. The method of Claim 16, wherein the method further comprises administering a therapeutically effective amount of a steroidal anti-inflammatory 25 drug.

18. The method of Claim 16, wherein the disease is a respiratory disease.

19. The method of Claim 18, wherein the disease is asthma or chronic 30 obstructive pulmonary disease.